Abstract: There are electrically conductive, electroactive functionalized conjugated polymers having formula (I): ##STR1## These electrically conducive, electroactive conjugated polymers may be covalently bonded to a first biological molecule or antiligand. Polymers bonded to a first biological molecule or antiligand may be used to form an electrode and may be used to assay for, detect and/or extract a second biological molecule or ligand.

Abstract: A chemical linking agent is formed of a di- or higher functional photoactivatable compound having at least one group that is charged to improve water solubility, and two or more photoreactive groups (preferably comprising an aryl ketone such as benzophenone) that allow use of the agent in aqueous systems. Charged groups include salts of organic acids such as sulfonate, phosphonate and carboxylate groups, onium groups such as quaternary ammonium, sulfonium and phosphonium groups, and protonated amines. A surface can be coated with a substance such as a synthetic polymer, carbohydrate, protein, lipid, nucleic acid, drug, vitamin, cofactor or dye by forming an aqueous solution of the linking agent and the substance, placing the solution in contact with the surface and activating the photoreactive groups of the linking agent to cross-link the substance to the surface.

Abstract: The present invention provides glycophospholipid and peptide-phospholipid conjugates comprising a phospholipid moiety and a saccharide or peptide moiety joined by an ether linkage comprising a secondary or tertiary amine. The conjugate structure of the invention comprises a flexible spacer arm between the phospholipid and saccharide or peptide moieties which, being variable in length, serves to optimize saccharide or peptide bioactivity. This invention further provides a method for the synthesis of such conjugates comprising the step of reductive amination. The method is efficient, economical and provides a high yield of product. Glycophospholipid and peptide-phospholipid conjugates of the invention can be incorporated and, optionally, chemically polymerized in self-assembling systems such as membranes, bilayers, films, liposomes and the like, and find utility diagnostically and therapeutically in medical and immuno-biological applications.

Abstract: The present invention provides methods for preparing, and compositions comprising, stabilized protein-polymer conjugates. More particularly, the present invention relates to the stabilization of individual subunits of multisubunit protein complexes by conjugation to polymers. Such conjugation acts to stabilize the individual subunit in its native conformation in liquid medium, which in turn acts to stabilize its biological activity.

Abstract: A viral agglutination test agent and a virus test kit containing the agent, which can directly and sensitively detect viruses in blood without using the anti-viral antibody formed in the blood of patients with viral infections.

Abstract: The present invention relates to the discovery of novel genes and proteins, which function in pathways involved in brain pathogenesis. In particular, the novel genes and proteins relate to inflammatory tissue responses caused by brain injuries such trauma, ischemia or autoimmune-inflammation or other diseases or processes related to neuroinflammation. The compounds disclosed in the present invention are useful as therapeutics, diagnostics and in screening assays.

Abstract: Macromolecules such as nucleic acids, proteins, lipids or polysaccharides are aligned on a support surface by passing the macromolecules through a meniscus of a solvent containing the macromolecules. The meniscus may be that of a solvent between two surfaces at an interface of the solvent with air. One end of a macromolecule is attached to one surface which may be a glass surface and another end is free. The meniscus is moved relative to the surface to which the end is attached such as by evaporating the solvent or by moving the surface. As the macromolecule passes through the meniscus, the macromolecule aligns on the surface perpendicular to the meniscus. This method may be used in assaying, measuring intramolecular distance and/or separating of macromolecules.

Abstract: Targeting molecules for use in delivering imaging agents to epithelial tissue are disclosed. Upon delivery, the imaging agent(s) may remain within an epithelial cell or may undergo transepithelial transport via transcytosis. The targeting molecules may be used, for example, for diagnostic techniques. The targeting molecule is a polypeptide, which may be produced by recombinant methods, that forms a closed covalent loop, contains at least three peptide domains having .beta.-sheet character which are separated by domains lacking .beta.-sheet character, specifically binds to a basolateral factor attached to a basolateral domain of an epithelial surface causing uptake of a linked imaging agent into cells of the epithelial surface, and is not a full length dimeric Iga. Preferably, the polypeptide is a J chain polypeptide, or a J chain polypeptide linked to an immunoglobulin heavy chain without an immunoglobulin light chain.

Abstract: A peptide comprising at least the following amino acid sequence:Lys Xa1 Phe Lys Arg Ile Val Xa2 Arg Ile Xaa Xa2 Phe Leu Arg Xa2 Leu Val (SEQ ID NO: 1)wherein, Xa1 represents a hydrophobic amino acid residue, each of Xa2 independently represents a hydrophilic amino acid residue, and Xaa represents an arbitrary amino acid residue;an antimicrobial agent, a medicine including a bacterial infection-treating agent and an endotoxin shock suppressant which each comprise the peptide as an active ingredient; and an endotoxin-removing agent comprising the peptide immobilized to an insoluble carrier.

Abstract: Proteinaceous polymers having repetitive units from naturally occurring structural proteins are employed as backbones for functionalities for crosslinking to provide strongly adherent tissue adhesives and sealants. Particularly, block copolymers having repeating units of elastin and fibroin are employed having lysine substitutions in spaced apart units, where the amino group can be crosslinked using difunctional crosslinking agents. The protein polymer contains at least 40 weight percent of repetitive units of 3 to 30 amino acids of at least one naturally occurring structural protein and at least two functional groups capable of reacting with a crosslinking agent to form a strongly adherent adhesive composition for bonding together separated tissue or for sealing tissue defects.

Abstract: The present invention relates to streptavidin proteins and peptides having a altered physical properties such as an increased stability or increased or decreased affinity for binding biotin. The invention also relates to methods for the detection, identification, separation and isolation of targets using streptavidin proteins or peptides. Streptavidin with increased or reduced affinity allows for the use of the streptavidin-biotin coupling systems for detection and isolation systems wherein it is necessary to remove of one or the other of the binding partners. Such systems are useful for the purification of functional proteins and viable cells. The invention also relates to nucleic acids which encode these streptavidin proteins and peptides and to recombinant cells such as bacteria, yeast and mammalian cells which contain these nucleic acids.

Abstract: Multiple layered functional thin films fixed on a solid support are provided which comprise multiple layers of functional molecules (such as enzymes and other proteins, pigments and dyes) admixed with polymer ions in combination with multiple layers of polymer ions without the functional molecules. The films are prepared by immersing a solid support having an electric charge in an admixed polymer ion-functional molecule solution having a net electric charge opposite to that of the solid support followed by immersing the solid support in a polymer ion solution having a net electric charge opposite to that of the admixed polymer ion-functional molecule solution, and repeating at least once the immersings of the solid support in the solutions.

Abstract: Methods are provided for forming a coating of an immobilized biomolecule on a surface of a medical device to impart improved biocompatibility for contacting tissue and bodily fluids. A 2-aminoalcohol moiety of a biomolecule is oxidized with a periodate to an aldehyde moiety which is reacted with an amine moiety on the surface of a medical device to form an imine moiety, and the imine moiety is reduced to form an amine linkage immobilizing a coating of the biomolecule on the surface. Conversely, the biomolecule can contain the amine moiety and the 2-aminoalcohol moiety can be on the surface. In another method, a biomolecule coating containing an amine moiety and a 2-aminoalcohol moiety is immobilized on the surface of a medical device, and the 2-aminoalcohol moiety is oxidized with a periodate to an aldehyde moiety which reacts with the amine moiety to crosslink the coating.

Abstract: Polynucleotide sequences are provided for the diagnosis of the presence of retroviral infection in a human host associated with lymphadenopathy syndrome and/or acquired immune deficiency syndrome, for expression of polypeptides and use of the polypeptides to prepare antibodies, where both the polypeptides and antibodies may be employed as diagnostic reagents or in therapy, e.g., vaccines and passive immunization. The sequences provide detection of the viral infectious agents associated with the indicated syndromes and can be used for expression of antigenic polypeptides.

Abstract: A protein such as an enzyme or antibody is immobilized by crosslinking crystals of the protein with a multifunctional crosslinking agent. The crosslinked protein crystals may be lyophilized for storage. A preferred protein is an enzyme such as thermolysin, elastase, asparaginase, lysozyme, lipase or urease. Crosslinked enzyme crystals preferably retain at least 91% activity after incubation for three hours in the presence of a concentration of Pronase.TM. that causes the soluble uncrosslinked form of the enzyme to lose at least 94% of its initial activity under the same conditions. A preferred enzyme:Pronase.TM. ratio is 40:1. Enzyme crystals that are crosslinked may be microcrystals having a cross-section of 10.sup.-1 mm or less.

Abstract: Proteins such as enzymes and antibodies are immobilized by crosslinking crystals of the proteins such as microcrystals having a cross-section of 10.sup.-1 mm or less with a multifunctional crosslinking agent. The crosslinked protein crystals may be lyophilized for storage. Crystals of an enzyme such as thermolysin, elastase, asparaginase, lysozyme, lipase or urease may be crosslinked to provide crosslinked enzyme crystals that retain at least 91% activity after incubation for three hours in the presence of a concentration of Pronase.TM. that causes the soluble uncrosslinked form of the enzyme to lose at least 94% of its initial activity under the same conditions. A preferred Pronase.TM.:enzyme ratio is 1:40.

Abstract: A method of determining the sequence of monomers which is a conformational equivalent of an epitope which is complementary to a particular paratope of an antibody of interest, the method comprising the steps of: 1. synthesizing a plurality of catamer preparations; each of said catamer preparations consisting of a plurality of catamers in which the composition at one or more designated positions in each catamer is known, and the composition at the remaining positions is randomly made up from members of a defined set of monomers; and said plurality of catamer preparations comprising preparations in which the composition at the designated positions is systematically varied to contain members from a defined set of monomers; 2. contacting each of the plurality of catamer preparations with the antibody of interest; and 3.

Abstract: Link protein and cartilage matrix protein, which are two major components of the extracellular cartilage matrix, have been found to bind to each other. Polypeptide fragments of cartilage matrix protein and link protein are produced. A recombinant fusion polypeptide is prepared containing a fragment of cartilage matrix protein that binds to link protein and a fragment of link protein that binds to cartilage matrix protein. The cartilage matrix protein fragment may bind to collagen and contain the CMP-1 or CMP-2 domain, and the link protein may bind to a complex of hyaluronic acid and proteoglycan. The fragments or fusion polypeptide can be administered for repair of diseased or injured cartilaginous and non-cartilaginous tissue by promoting binding of a complex of proteoglycan and hyaluronic acid to collagen. The fragments or fusion polypeptide can be anchored to the surface of a prosthetic device, implant or tissue graft to promote adherence of tissue and biocompatibility.

Abstract: A method for the detection of a polynucleotide target sequence is described. The method involves the formation of a covalent or non-covalent bonded pair of nucleotide sequences formed in response to a target polynucleotide sequence, adding nucleotide sequence specific binding proteins each capable of binding one member of the pair of nucleotide sequences, and detecting the specific binding proteins complexed to the pair of nucleotide sequences.

Abstract: The invention provides methods for the purification to homogeneity of pancreatic cholesterol esterase in useful quantities from a variety of mammalian species. The gene for a mammalian pancreatic cholesterol esterase has been cloned and sequenced, and is useful for expressing cholesterol esterase in a transformed eukaryotic or prokaryotic cell culture. Thus, methods according to the invention enable the production of large quantities of pancreatic cholesterol esterase for the screening of inhibitors, the production of antibodies, and for commercial purposes related to the alteration of cholesterol/cholesterol ester composition of materials containing free or esterified cholesterol.

Abstract: A protein such as an enzyme or antibody is immobilized by crosslinking crystals of the protein with a multifunctional crosslinking agent. The crosslinked protein crystals may be lyophilized for storage. A preferred protein is an enzyme such as thermolysin, elastase, asparaginase, lysozyme, lipase or urease. Crosslinked enzyme crystals preferably retain at least 91% activity after incubation for three hours in the presence of a concentration of Pronase.TM. that causes the soluble uncrosslinked form of the enzyme to lose at least 94% of its initial activity under the same conditions. A preferred enzyme:Pronase.TM. ratio is 1:40. Enzyme crystals that are crosslinked may be microcrystals having a cross-section of 10.sup.-1 mm or less.

Abstract: The invention provides an extracellular matrix for enhancing wound healing. The extracellular matrix comprises a recombinant fibronectin protein and a backbone matrix, wherein the recombinant fibronectin protein comprises peptides from two or more fibronectin domains. The extracellular matrix facilitates wound healing by providing hemostasis and, in addition, an environment that intrinsically recruits new tissue cells to the wound site. The extracellular matrix according to the subject invention is thus used in a method for enhancing wound healing. The method comprises applying the extracellular matrix to the wound.

Abstract: Crosslinking to prepare fixed tissue or to crosslink modification molecules to tissue is carried out with an apparatus and method wherein diffusion through a semipermeable is used to provide a selected molecular weight distribution of oligomers of a crosslinking compound that polymerizes spontaneously in solution to produce oligomers such as glutaraldehyde. The membrane has a molecular weight exclusion limit that prevents oligomers above a certain molecular weight from passing. Tissues treated with the size selected oligomers have improved properties. In the apparatus, the membrane separates a solution containing the crosslinking compound and oligomers from a solution in which crosslinking is carried out. The selected molecular weight distribution of oligomers diffuses through the membrane into the solution where tissue is crosslinked or a beneficial molecule is crosslinked to tissue.

Abstract: An immunogen comprising a residue of a histamine-releasing peptide comprising a cationic N-terminal head and a hydrophobic C-terminal tail, together with a residue capable of eliciting antibodies against said peptide whilst inhibiting histamine release by said peptide is useful in anti-allergy treatment. Preferably the histamine-releasing peptide is of formula:Lys-Thr-Lys-Gly-Ser-Gly-Phe-Phe-Val-Phe,optionally amidated at the C terminal.Antibodies to the histamine-releasing peptide are useful for passive immunisation.

Abstract: The invention provides an immobilised oligopeptide for detecting protein prenylation consisting of an oligopeptide containing the amino acid sequence Xad-Xac-Xab-Xaa-OH at its carboxyl-terminus, at least one of Xaa, Xab, Xac and Xad representing cysteine (Cys), said sequence being capable of acting as a substrate for a prenyl transferase catalysing protein prenylation, and being bonded to a solid carrier, preferably at its amino-terminus. The invention further provides a kit for detecting protein prenylation comprising a first immobilised oligopeptide as mentioned above and a second immobilised oligopeptide which differs from said first immobilised oligopeptide in that the cysteine residue is substituted by another amino acid. Also provided are antibodies against the prenylated oligopeptides.

Abstract: A method for screening compounds for antimicrobial activity is described that utilizes bacterial protein-protein binding in vitro. The method may be performed using immobilized elements and the immobilization may be carried out using a variety of immobilization means (e.g., columns, beads, adsorbents, nitrocellulose paper, etc.) in order to screen large libraries of compounds.

Abstract: Methods are provided for forming a coating of an immobilized biomolecule on a surface of a medical device to impart improved biocompatibility for contacting tissue and bodily fluids. A 2-aminoalcohol moiety of a biomolecule is oxidized with a periodate to an aldehyde moiety which is reacted with an amine moiety on the surface of a medical device to form an imine moiety, and the imine moiety is reduced to form an amine linkage immobilizing a coating of the biomolecule on the surface. In another method, a biomolecule coating containing an amine moiety and a 2-aminoalcohol moiety is immobilized on the surface of a medical device, the 2-aminoalcohol moiety is oxidized with a periodate to an aldehyde moiety which is reacted with the amine moiety to form an imine moiety, and the imine moiety is reduced to form a secondary amine and crosslink the coating.

Abstract: Compositions and methods for avidin immobilized on an inert support material, e.g., agarose, are disclosed. The compositions have high activity levels of avidin and may further include a bulking agent, e.g., maltose, and a protectant to maintain the stability and integrity of the avidin agarose during lyophilization and terminal sterilization processes. A dry composition, and wet compositions such as a gel, slurry or suspension having avidin immobilized on an inert support material are disclosed. The dry composition has at least 1000 biotin binding units of activity, and the wet compositions have at least 50 units of binding activity. These compositions have applicability in any instance where avidin agarose and/or the avidin/biotin technology are useful. In particular, the present compositions are useful in an enzyme capture system to prepare fibrin monomer useful for fibrin sealants.

Abstract: Polyionic derivatives of cyclodextrins and methods for preparing these derivatives are provided in which a polyionic derivative of cyclodextrin is combined with a growth factor, preferably a heparin binding growth factor. These compositions are of low solubility and are applied directly to the location of a wound. By virtue of the low solubility, the compositions remain in place at the site of application and slowly release growth factor. In an alternative embodiment, the cyclodextrin derivatives are administered in the absence of growth factor and are used to absorb growth factor present in the body at the location of the wound in order to prevent overstimulation of the wound response.

Abstract: Biotinylation peptides can be fused to other peptides or proteins of interest using recombinant DNA techniques to provide efficient methods for biotinylating the resulting fusion proteins in vivo or in vitro.

Abstract: Methods are provided for forming a coating of an immobilized biomolecule on a surface of a medical device to impart improved biocompatibility for contacting tissue and bodily fluids. A biomolecule having a negatively charged moiety is combined with a medical device surface having a positively charged guanidino moiety to form an ionic bond immobilizing a coating of the biomolecule on the surface. In another method, the medical device surface contains an amine moiety that is combined with a guanidino forming agent to form a positively charged guanidino moiety that is combined with the negatively charged moiety to form the ionic bond. In a further embodiment, the medical device surface contains a negatively charged moiety, and a biomolecule containing an amine moiety is combined with a guanidino forming agent to form a positively charged guanidino moiety that is combined with the negatively charged moiety to form the ionic bond.

Abstract: Methods are provided for forming a coating of an immobilized biomolecule on a surface of a medical device to impart improved biocompatibility for contacting tissue and bodily fluids. A biomolecule such as a glycoprotein having an unsubstituted amide moiety is combined with an amine forming agent to form an amine-functional biomolecule. The amine-functional biomolecule is combined with a medical device surface having a chemical moiety such as aldehyde, epoxide, isocyanate, 1,2-dicarbonyl, phosphate, sulphate or carboxylate to form a chemical bond immobilizing the biomolecule on the surface. The chemical bond may be combined with a reducing agent or a stabilizing agent. The aldehyde moiety may be formed by combining a periodate with a 2-aminoalcohol moiety or a 1,2-dihydroxy moiety. Alternatively, an amine-functional medical device surface is combined with a biomolecule having a chemical moiety that reacts with an amine moiety.

Abstract: Compounds and methods are provided for diagnosing Leishmania infection. Disclosed compounds include polypeptides that contain at least an epitope of the Leishmania chagasi acidic ribosomal antigen LcP0, or a variant thereof. Such compounds are useful in a variety of immunoassays for detecting Leishmania infection and for identifying individuals with asymptomatic infections that are likely to progress to acute visceral leishmaniasis. The polypeptide compounds are further useful in vaccines and pharmaceutical compositions for preventing leishmaniasis.

Abstract: Antivenoms to snake, spider, scorpion and jelly fish venoms are produced for treatment of humans and animals, and for analytical use. Polyvalent antivenoms are produced containing immunoglobulin which is greater than fifty percent venom reactive. Purified polyvalent antivenom is derived from a first polyvalent antivenom having two or more monovalent subpopulations, and purified such that greater than fifty percent of the monovalent subpopulations are recovered by weight. The antivenoms can be horse or avian such as chicken antivenom. Chicken antivenom is obtained using a whole venom that is not glutaraldehyde pretreated, and the antivenom contains yolk immunoglobulin. Antivenoms are purified with an antigen matrix containing a single whole venom or a plurality of whole venoms covalently attached to an insoluble support such as aldehyde-activated agarose. Preferably, the whole venoms forming the plurality of whole venoms are selected from the four whole venoms of C. atrox, B. atrox, C. adamanteus and C.

Abstract: Polyionic derivatives of cyclodextrins and methods for preparing these derivatives are provided in which a polyionic derivative of cyclodextrin is combined with a growth factor, preferably a heparin binding growth factor. These compositions are of low solubility and are applied directly to the location of a wound. By virtue of the low solubility, the compositions remain in place at the site of application and slowly release growth factor. In an alternative embodiment, the cyclodextrin derivatives are administered in the absence of growth factor and are used to absorb growth factor present in the body at the location of the wound in order to prevent overstimulation of the wound response.

Abstract: The present invention is directed to isolated amyloid precursor-like proteins (APLPs), nucleotide sequences coding for and regulating expression of these protein, antibodies directed against these proteins, and recombinant vectors and host cells containing the genetic sequences coding for and regulating the expression of these protein sequences. The invention is also directed to isolated genomic. DNA, cDNA anti-sense RNA, and RNA containing the protein sequence. Antibodies can be used to detect an APLP in biological specimens, including, for example, fluid, serum or tissue samples. APLP1 and APLP2 are candidate genes for late onset familial Alzheimer's disease.

Abstract: The present invention relates generally to intracellular receptor recognition proteins or factors, termed Signal Transducers and Activators of Transcription (STAT), to methods and compositions utilizing such factors, and to the antibodies reactive toward them, in assays and for diagnosing, preventing and/or treating cellular debilitation, derangement or dysfunction. More particularly, the present invention relates to particular functional domains of molecules that exhibit both receptor recognition and message delivery via DNA binding in receptor-ligand specific manner, i.e., that directly participate both in the interaction with the ligand-bound receptor at the cell surface and in the activity of transcription in the nucleus as a DNA binding protein. The invention likewise relates to the antibodies and other entities that are specific to the functional domain of a STAT protein and that would thereby selectively modulate its activity.

Abstract: Link protein and cartilage matrix protein, which are two major components of the extracellular cartilage matrix, have been found to bind to each other. Cartilaginous tissue is attached to a surface by anchoring on the surface a fragment of cartilage matrix protein capable of binding to link protein or to collagen and link protein, or a fragment of link protein capable of binding to cartilage matrix protein or to cartilage matrix protein and a complex or proteoglycan and hyaluronic acid, and contacting the surface with cartilaginous tissue. Cartilage matrix protein is attached to a surface by anchoring on the surface a fragment of link protein capable of binding to cartilage matrix protein, and contacting the surface with cartilage matrix protein. Link protein is attached to a surface by anchoring on the surface a fragment of cartilage matrix protein capable of binding to link protein, and contacting the surface with link protein.

Abstract: An oligopeptide having an amino acid sequenceGlu Pro Gly Asn Ser Glu Ile Leu Pro Thr Leu Lysand variants thereof; immunogenic conjugates obtained therefrom; antibodies produced by means of said conjugates and which specifically recognize human plasma glutathione peroxidase (pl.GPx); methods for assaying human plasma glutathione peroxidase (pl.GPx); and assaying kits. The invention is useful in the medical diagnosis, treatment and monitoring of pathologic conditions induced by a variation in pl.GPx, for example, hepatic tumors, acute rejection of renal or hepatic graft, renal insufficiency and selenium deficiency.

Abstract: A variant or so-called "escape mutant" HBsAg protein or fragment thereof displaying the antigenicity of hepatitis B virus surface antigen is disclosed, in which the mutant protein or fragment thereof (mHBsAg) comprises a modified `a` determinant in which at least two amino acids are inserted downstream of position 122 of the wild type HBsAg sequence. A vaccine comprising the mHBsAg is provided, as is a kit for diagnostic in vitro detection of anti-mHBsAg antibodies and an antibody preparation comprising anti-mHBsAg antibodies.

Abstract: A method for making a medical device having a biomolecule immobilized on a substrate surface is provided. The method includes: providing an immobilized biomolecule comprising a biomolecule covalently attached to a support material; attaching a photoreactive crosslinking agent to the immobilized biomolecule to form a photoreactive analog of the biomolecule; and removing the photoreactive analog of the biomolecule from the support material. The photoreactive analog of the biomolecule can then be attached to a substrate surface, such as a biomaterial that forms part of a medical device. The immobilized biomolecule may contain a peptide having an N.sup..alpha. -terminus. The photoreactive crosslinking agent is attached to the peptide at the N.sup..alpha. -terminus to form the photoreactive analog of the biomolecule. The peptide can be an adhesion peptide containing the sequence Trp-Gln-Pro-Pro-Arg-Ala-Arg-Ile. Attachment of the peptide to a substrate surface promotes cell adhesion to the surface.

Abstract: A new class of phenylboronic acid complexing reagents are provided capable of binding with phenylboronic acids is disclosed having one of the following structures: ##STR1## wherein group X is selected from H, CH.sub.3 and C.sub.6 H.sub.5 ; and groups Y and Z are selected from O and CH.sub.2 ; group Q is a spacer which is from 2 to 12 carbon equivalents in length, and which may contain intermediate amide and/or ether functionalities; and group R is a reactive electrophilic moiety suitable for conjugation of a phenylboronic acid complexing reagent with a biological macromolecular species, low molecular weight species or solid phase support having a reactive pendant nucleophilic moiety. The phenylboronic acid complexing reagents are utilized in conjunction with phenylboronic acid reagents to facilitate chemical conjugation without the use of intermediary biological macromolecules. The method of making and using such reagents is also disclosed.

Abstract: A trifunctional conjugate is provided having three chemical moieties attached through a spacer moeity. At least two of the chemical moieties are relatively small molecules, usually less than about 7,000 Daltons in size. The spacer moiety is selected to impart certain steric properties to the conjugate. In one embodiment, the binding of a macromolecular specific binding partner to one of the chemical moieties sterically inhibits the binding of a different macromolecule to another chemical moiety. In another embodiment, the binding of a first chemical moiety to a macromolecule restricts the subsequent binding of a second chemical moiety to a proximate location on the same macromolecule. The three chemical moieties are preferably a nitrophenylazido residue, a phenyl boronic acid residue, and a solid support or a label such as biotin. The spacer is preferably cysteine, lysine, glutamic acid, pyroglutamic acid, S-acetylmercaptosuccinic anhydride or .omega.-carbobenzoxylysine.

Abstract: A protein such as an enzyme or antibody is immobilized by crosslinking crystals of the protein with a multifunctional crosslinking agent. The crosslinked protein crystals may be lyophilized for storage. A preferred protein is an enzyme such as thermolysin, elastase, asparaginase, lysozyme, lipase or urease. Crosslinked enzyme crystals preferably retain at least 91% activity after incubation for three hours in the presence of a concentration of Pronase.TM. that causes the soluble uncrosslinked form of the enzyme to lose at least 94% of its initial activity under the same conditions. A preferred enzyme:Pronase.TM. ratio is 1:40. Enzyme crystals that are crosslinked may be microcrystals having a cross-section of 10.sup.-1 mm or less.

Abstract: Macromolecules such as nucleic acid strands are aligned, adhered and stretched on a support surface by passing the strands through a meniscus of a solvent containing the strands. The meniscus may be that of a solvent between two surfaces at an interface of the solvent with air. One end of a nucleic acid strand is attached to one surface which may be a glass surface and another end is free. The meniscus is moved relative to the surface to which the end is attached such as by evaporating the solvent or by moving the surface. As the nucleic acid strand passes through the meniscus, the strand elongates and aligns perpendicular to the meniscus on the surface. This method may be used in assaying, measuring intramolecular distance of and/or separating macromolecules.

Abstract: A method is provided for immobilizing a biomolecule coating on the surface of a medical device to obtain improved biocompatibility characteristics for contacting with tissue or body fluids such as blood. The method includes oxidizing a 2-aminoalcohol moiety of a material disposed on the surface with a periodate to form an aldehyde moiety, combining the aldehyde moiety with an amine moiety of a material to bond the aldehyde moiety to the amine moiety through an imine moiety and reacting the imine moiety with a reducing agent to form the coating immobilized on the surface by an amine linkage.

Abstract: Proteinaceous polymers having repetitive units from naturally occurring structural proteins are employed as backbones for functionalities for crosslinking to provide strongly adherent tissue adhesives and sealants. Particularly, block copolymers having repeating units of elastin and fibroin are employed having lysine substitutions in spaced apart units, where the amino group can be crosslinked using difunctional crosslinking agents. The protein polymer contains at least 40 weight percent of repetitive units of 3 to 15 amino acids of at least one naturally occurring protein and in at least two units an amino acid is substituted by an amino acid containing a functional group capable of reacting with a crosslinking agent to form a strongly adherent adhesive composition for bonding together separated tissue or for sealing tissue defects.

Abstract: A method is provided for producing a sterile and pyrogen-free column containing coupled protein for use in removing a predetermined substance from the blood of a human subject. The method abrogates sterilization of the finished protein-containing product by providing sterile and pyrogen-free raw materials at each production step. The method provides a pathogen-free, purified solution of protein which binds to a predetermined substance in human blood such as LDL or immunoglobulin. Typically, the protein is anti-human LDL immunoglobulin or anti-human Ig immunoglobulin. The method also provides a sterile and pyrogen-free column matrix material such as an agarose which is chemically activated, either using CNBr and triethylamine or using 1,1'-carbonyldiimidazole.

Abstract: A protein such as an enzyme or antibody is immobilized by crosslinking crystals of the protein with a multifunctional crosslinking agent such as glutaraldehyde, and if desired lyophilizing the crosslinked crystals for storage. Crosslinking of the protein crystals provides stabilization for use under harsh conditions and for lyophilizing. The crystals crosslinked may be microcrystals having a cross-section of 10.sub.-1 mm or less. Crosslinked thermolysin, esterase, elastase, asparaginase and lysozyme crystals and crosslinked crystals of lipase from Geotrichum candidum and Candida cylindracea and of porcine origin can be used to convert a substrate to a product. Crosslinked thermolysin crystals are prepared that retain at least 96% of their initial activity after incubation for 4 days in the presence of a concentration of Pronase.TM. such as a thermolysin:Pronase.TM.

Abstract: A method for the separation of at least one specific binding entity from a mixture of binding entities by the steps of contacting a mixture of binding entities with immobilized peptides in which the peptides specifically bind to the specific binding entities and the peptides contain an amino acid which facilitates removal of the binding entities from the peptides, and separating the immobilized peptide/specific binding entity complexes from the mixture of binding entities. A change in incubation conditions facilitates removal of the binding entities from the immobilized peptides.