Abstract: A method is disclosed for obtaining highly specific and tailored ligands suitable for purifying a particular product target or for eliminating particular target impurities in a feed stream. Engineered affinity ligands according to the invention will bind a target with high specificity at a preselected binding condition and release the target at a preselected elution condition. The ligands are isolated by contacting a target with a multiplicity of polypeptides derived through variegation of the structure of a candidate binding domain, the variants (or analogues) including polypeptides favoring binding to the target under desired binding conditions and release from the target under elution conditions, where the binding and elution conditions differ according to one or more parameters, such as pH, temperature, concentration of salt or volume % of an organic solvent.

Abstract: The present invention relates to the C3b/C4b receptor (CR1) gene and its encoded protein. The invention also relates to CR1 nucleic acid sequences and fragments thereof comprising 70 nucleotides and their encoded peptides or proteins comprising 24 amino acids. The invention further provides for the expression of the CR1 protein and fragments thereof. The genes and proteins of the invention have uses in diagnosis and therapy of disorders involving complement activity, and various immune system or inflammatory disorders. In specific embodiments of the present invention detailed in the examples sections infra, the cloning, nucleotide sequence, and deduced amino acid sequence of a full-length CR1 cDNA and fragments thereof are described. The expression of the CR1 protein and fragments thereof is also described. Also described is the expression of a secreted CR1 molecule lacking a transmembrane region.

Abstract: Novel compositions and methods useful for the generation of nucleic acids from a ribonucleic acid template and further nucleic acid replication are disclosed. It is shown that the generation and amplification of nucleic acids by methods that utilize two or more different polymerases, such as reverse transcriptase-polymerase chain reaction (RT-PCR), are dramatically more sensitive and efficient in the presence of a homopolymeric nucleic acid. Homopolymeric nucleic acids have been found to reduce or negate the inhibitory effect reverse transcriptases have on DNA polymerase activity. It is demonstrated that this inhibition-relieving effect of homopolymeric nucleic acids is general in nature; independent of the chemical species of homopolymer used, or the chemical composition of the polymerization reaction mixture.

Abstract: Methods for detecting human parvovirus B19 in and removing it from biological samples such as blood are disclosed, together with reagents suitable for the purpose comprising binding moieties that recognize human parvovirus B19 and/or B19-like polypeptide and form a binding complex therewith. Preferred polypeptide binding moieties are particularly disclosed.

Abstract: Splice variants of known TAP1 and TAP2 proteins, which are involved in translocation of antigen peptides into the endoplasmic reticulum for complexing with MHC class I molecules and eventual display on the cell surface, are disclosed. A fully sequenced and characterized splice variant of TAP2, designated TAP2iso, is shown to form functional heterodimers with TAP1 and to exhibit a peptide specificity that differs from previously studied TAP1/TAP2 transporter proteins. The discovery of splice variant TAP subunits alters the prior theory of immune response and introduces a mechanism for diversification of antigen display to the CD8-positive T cells of the immune system. Methods for diagnosis and treatment of diseases or conditions associated with abnormal TAP splice variant expression, or of expanding the reptertoire of antigen peptides to which an individual's immune system is capable of responding, are also disclosed.

Abstract: A plasmid-based vaccine is provided herein based on the combination of DNA segments coding for one or more B cell epitopes of cholesteryl ester transfer protein (CETP) and one or more broad range helper T cell epitopes. Administration of the plasmids as a vaccine to a vertebrate subject provides an immune response to the subject's endogenous CETP and modulation of CETP activity, leading to prevention or reversal of various manifestations of heart disease. The vaccines provide an advantageous strategy for the prevention or treatment of atherosclerosis.

Abstract: A method is disclosed for the rapid diagnosis of disorders characterized by an ischemic event, such as stroke, transient ischemic attack, head trauma, myocardial infarction or other insults resulting in interrupted cranial blood flow. The method involves detection of the presence of the gamma isoform of protein kinase C (PKCg) in peripheral blood, which signals damage to central nervous system tissue and at least transient breakdown of the blood brain barrier. The assay may be performed, e.g., by emergency medical personnel, in a time frame that allows treatment of the patient before permanent damage to the central nervous system occurs.

Abstract: The use of multi-well blocks with more than 10 wells is suggested for breaking down biological samples in a ball mill, wherein the process which is to be carried out provides that the biological sample, disposed in a multi-well block with more than 10 wells, is broken down in a ball mill and that following this, proteins and/or nucleic acids are isolated. With the ball mill necessary for this task, with milling containers fixed thereto in a holder (10), the milling containers are designed as uniform well blocks (16) and can be fixed in a form-locking manner into the adapted, twist-preventing holder (10).

Abstract: The present invention relates to a method of producing covalently closed circular (ccc) DNA from a cultured cell or unicellular organism, preferably a bacterium, essentially free of genomic DNA. The method of the invention comprises the steps of precipitating a cleared lysate with an alcohol, washing and resuspension of the precipitate, digestion of the genomic DNA with a nuclease that cleaves linear DNA or circular DNA that includes a nick or a free 3′ or 5′ end but not ccc DNA, and finally, separation of the purified ccc DNA from the remainder of the product of the digestion step by contacting the product with an ion exchange material. ccc DNA obtained by the method, pharmaceutical compositions comprising the ccc DNA, and a kit for carrying out the method of the invention are also contemplated.

Abstract: Methods for detecting human parvovirus B19 in and removing it from biological samples such as blood are disclosed, together with reagents suitable for the purpose comprising binding moieties that recognize human parvovirus B19 and/or B19-like polypeptide and form a binding complex therewith. Preferred polypeptide binding moieties are particularly disclosed.

Abstract: Methods for detecting human factor VIII or factor VIII-like polypeptides in and isolating it from solutions such as blood or conditioned media are disclosed, together with reagents suitable for the purpose comprising binding moieties that recognize human factor VIII and/or a factor VIII-like polypeptide and form a binding complex therewith. Preferred polypeptide binding moieties are particularly disclosed.

Abstract: The present invention provides compositions comprising at least one complement moiety and at least one carbohydrate moiety, and methods of producing such compositions. In particular, the compositions of the invention comprise complement proteins related to the complement receptor type 1, and further comprise ligands for intercellular molecules, such as selectins. In a preferred embodiment, the compositions comprise a complement-related protein in combination with the Lewis X antigen or the sialyl Lewis X antigen. The compositions of the invention have use in the diagnosis or therapy of disorders involving complement activity and inflammation. Pharmaceutical compositions are also provided for treating or reducing inflammation mediated by inappropriate complement activity and intercellular adhesion.

Abstract: A method for the amplification of a target nucleic acid is disclosed comprising the steps of reacting a nucleic acid with an amplification reaction mixture and a modified thermostable enzyme, wherein said modified thermostable polymerase is prepared by a reaction of a mixture of a thermostable polymerase and a chemical modifying reagent. The chemical modification reagent is an aldehyde, preferably formaldehyde. Essentially complete inactivation of the enzyme at ambient temperatures is achieved, with recovery of enzymatic activity at temperatures above 50° C.

Abstract: The present invention relates to methods of immunization of aquaculture species by introducing DNA expression systems into the aquaculture species. Such DNA expression systems preferably include DNA sequences encoding polypeptides of pathogens of species of aquaculture. The present invention also relates to methods of administration of DNA expression systems into aquaculture. Such methods include injection, spray, and immersion techniques. The methods of this invention are useful for prophylactic vaccination or therapeutic immunization of fin-fish, shellfish, or other aquatic animals against infectious diseases.

Abstract: A method is disclosed for treating diseases or disorders involving complement by pulmonary administration of complement inhibitory proteins such as soluble complement receptor type 1 (sCR1). The present invention relates to the direct treatment of certain complement related disorders by administering complement inhibitory proteins via the pulmonary route, in particular, by direct delivery to the lungs by aerosolization of a complement inhibitory protein and subsequent inhalation.

Abstract: This invention provides: novel proteins, which are homologous to the first Kunitz domain (K1) of lipoprotein-associated coagulation inhibitor (LACI), and which are capable of inhibiting plasmin; uses of such novel proteins in therapeutic, diagnostic, and clinical methods; and polynucleotides that encode such novel proteins.

Abstract: Several innovative systems for an aircraft, and aircraft incorporating them, are disclosed. Features include inboard-mounted engine(s) with a belt drive system for turning wing-situated propellers; compound landing gear integrating ski, pontoon and wheel subcomponents; pivotal mounting armatures for landing gear and/or propellers which provide a plurality of possible landing gear and/or propeller configurations; and a compound wing structure featuring extendable wing panels that permit the wing span of the aircraft to be nearly doubled while in flight. Aircraft incorporating such features will enjoy several safety advantages over conventional multi-engine aircraft and will be capable of modifications during flight which permit landings on any of snow, hard surfaces (runways) and water.

Abstract: A method is disclosed for obtaining affinity ligands for isolating tissue-type plasminogen activator (tPA). Ligands binding tPA with high specificity at pH 7 and releasing tPA at pH 5 or lower are disclosed.

Abstract: This invention provides oligonucleotides encoding novel Kunitz domain polypeptides capable of binding plasmin, and vectors and transformed host cells containing these polynucleotides.

Abstract: Provided are methods of making preparations of recombinant soluble Complement Receptor type 1 (sCR1) defined with respect to the distribution of sCR1 glycoforms and therapeutic methods using them. The preparations are suitable for treatment of allograft or xenograft rejection, diseases involving inflammation or inappropriate complement activation, and thrombotic or shock state conditions. Preferred methods employ sCR1 glycoforms that are sialylated, have a pI of .ltoreq.5.1, or have a sialic acid: mannose molar ratio of .gtoreq.0.25.