Abstract: The present invention concerns a method for the determination of allele-specific peptide motifs on molecules of the major histocompatibility complex (MHC) of classes I and II as well as the peptide motifs which are obtainable by the method according to the invention. In addition the use of the peptide motifs according to the invention for the production of a diagnostic or therapeutic agent is disclosed.

Abstract: A diagnostic method and screening test for atherosclerosis and analogous diseases involving activated phagocytes and/or inflammation is provided which comprises determining the presence of p-hydroxyphenylacetaldehyde-lysine in a test sample of a body fluid or tissue at a level which is elevated relative to the level in a normal patient.

Abstract: A solid support having surface-attached carboxylic acid groups is reacted with an isoxazolium salt to form an activated support having enol ester and sulfonate groups. After washing to remove residual isoxazolium salt and base, the activated support is dried. A polypeptide is covalently or non-covalently bound to the support. The resultant immobilized polypeptide can be conveniently sequenced by N- and C-terminal sequencing methods. The support can be a polyvinylidene difluoride membrane or a glass fiber membrane having surface attached carboxylic acid groups, and the enol and sulfonate groups are provided by reacting the support with 2-ethyl-5'-phenylisoxazolium sulfonate. The dry support can be stored for at least about 3 months to achieve a peptide immobilization yield that is substantially the same as the yield obtained in the absence of drying.

Abstract: A first reactant is immobilized i.e. in a porous matrix (50), adjacent a sample electrode (46) within a reaction chamber. Energizing of the electrode (46) electrophoretically attracts a mobile second reactant and/or electrolytically induces appropriate reaction conditions to enhance reaction of the first and second reactants. Polarity reversals between the sample electrode (46) and remote electrodes (38), (42), (44) cause unreacted second reactant and/or by-products to migrate away from the immobilized first reactant. The techniques are useful for sequential chemical reactions such as sequencing or construction of proteins, polysaccharides and nucleic acids where cyclical additions and removals of reactants are required. The techniques are amenable to automated micro and nano scale construction and operation and allow direct electrophoretic (38) interfacing with chromatographic, HPCE and mass spectrophotometric equipment.

Abstract: A method and device for forming large arrays of polymers on a substrate (401). According to a preferred aspect of the invention, the substrate is contacted by a channel block (407) having channels (409) therein. Selected reagents are delivered through the channels, the substrate is rotated by a rotating stage (403), and the process is repeated to form arrays of polymers on the substrate. The method may be combined with light-directed methodolgies.

Abstract: A method of forming a thiohydantoin from an N-protected amino acid is described. The method employs a phosphate compound selected from the group consisting of(R.sub.1 O)(R.sub.2 O)P(.dbd.O)X and(R.sub.1 O)(R.sub.2 O)P(.dbd.O)--O--P(.dbd.O)(OR.sub.3)(OR.sub.4)to form acylphosphate moieties from the carboxyl groups of internal aspartic acid and glutamic acid residues and an acylphosphate moiety at a C-terminal carboxyl. The later acylphosphate, unlike the internal acylphosphates, spontaneously cyclizes to an oxazolone, which is less reactive with nucleophilic reagents. R.sub.1 and R.sub.2 are each alkyl, aryl, or alkaryl groups which are the same or different and which may be covalently linked to each other; R.sub.3 and R.sub.4 are each alkyl, aryl, or alkaryl groups which are the same or different and which may be covalently linked to each other; and X is a leaving group, such as chlorine or bromine, which is substantially unreactive towards thiohydantoins.

Abstract: The binding sites of binding proteins and their binding partners are characterized, at the individual amino acid level, by a combination of tritium exchange labeling and sequential degradation and analysis of tritiated fragments under slowed exchange conditions.

Abstract: A machine for synthesizing oligonucleotides has individual pumping modules for connection to each of a number of different monomers and to other fluids used in the synthesizing process. Each pumping module includes a pump and a three-port/three-way valve with the first inlet port connected to receive the fluid, and the second port connected to receiving a flushing agent, such as acetonitrile. A valving arrangement downstream from the pumping modules selects a monomer. The selected monomer and other fluids pass through flow sensors which provides signals to a controller, which uses these signals to regulate the pumping of the pumps. At least some of the valves allow multiple inlet ports to be kept open at the same time, thus allowing certain of the liquids to be mixed within the valves. The various pumps, valves and sensors are mounted to the exterior of a cabinet and arranged such that the liquid handling portions are exterior to the cabinet, while electrical portions are within the cabinet.

Abstract: The present invention provides for an efficient and novel method for the C-terminal sequencing of proteins or peptides by use of acetyl chloride or phosphoryl chloride by reaction with a suitable isothiocyanate for derivitazation of the carboxy terminus to a thiohydantoin amino acid derivative, under acidic conditions. Cleavage of the derivatized thiohydantoin amino acid may occur by use of thiocyanic acid and acetic acid in water and also by the novel means using a buffer and a potassium or sodium thiocyanate or potassium or sodium dithionite reagent. The present invention also provides for an novel and efficient means for the C-terminal sequencing of proteins or peptides by a two or three step process which comprises first reacting the peptide or protein with an acid chloride reagent, such as acetyl chloride, or phosphoryl chloride.

Abstract: The present invention is directed to derivatives of aroyl-2-quinoline-carboxaldehyde and their use in detection and quantification of minute amounts of primary amines.

Abstract: The present invention provides for novel reagents whose fluorescence increases in the presence of particular proteases. The reagents comprise a characteristically folded peptide backbone each end of which is conjugated to a fluorophore. When the folded peptide is cleaved, as by digestion with a protease, the fluorophores provide a high intensity fluorescent signal at a visible wavelength. Because of their high fluorescence signal in the visible wavelengths, these protease indicators are particularly well suited for detection of protease activity in biological samples, in particular in frozen tissue sections. Thus this invention also provides for methods of detecting protease activity in situ in frozen sections.

Abstract: This invention relates to methods and apparatus for carrying out chemical reactions between a plurality of reactants and in particular it is amenable to micro or nano scale operation and to the sequential chemical reactions required during such processes as construction or sequencing of proteins, oligonucleotides and polysaccharides. The present invention further relates to a capillary liquid chromatography system for high-sensitivity component separation and microsequencing for use in association with the methods and apparatus herein described.

Abstract: A solid support and method for immobilization and sequence analysis of polypeptides. In one aspect, the invention is directed to a method for immobilizing a polypeptide on a solid support. In the method, a solid support having surface-attached carboxylic acid groups is reacted with an isoxazolium salt to form an activated support. After the activated support has been washed to remove residual isoxazolium salt and base, the support is dried. The dried support is then contacted with a polypeptide under conditions effective to bind the polypeptide covalently to the support. The immobilized polypeptide can be conveniently sequenced by N- and C-terminal sequencing methods.

Abstract: The subject invention provides a method for determining the most stable tertiary structure of a protein having a known primary structure which comprises the steps of (a) producing a reduced representation of the protein by assigning to the protein (i) all secondary structural motifs present therein and (ii) all .phi. and .PHI. dihedral angles for the amino acid residues present therein; (b) determining which conformations of the reduced representation are physically permissible, so as to determine which conformations of the protein are physically permissible; and (c) determining which of the physically permissible conformations of the protein possesses the lowest free energy, so as to thereby determine the most stable tertiary structure of the protein.

Abstract: The present invention further provides novel synthetic control peptides containing from about 3 to 100 natural amino acid residues that are designed for use in monitoring the proper operation of amino acid sequencers and to monitor peptide or protein cleavage reactions. The control peptide, or mixture of control peptides, are designed to obtain data for many or all common, uncommon and difficult to measure amino acids within 15 sequencer cycles and to provide cleavage sites for at least 4 different amino acid cleavage reactants.

Abstract: A universal standard chemical reagent for quantitative visual and spectrometric analysis of compounds having reactive functional groups, including mixtures and homologs of said compounds, said reagent comprising a compound of the general formula Q-B-f, wherein: Q stands for an organic moiety which can be measured quantitatively, visually by color, spectroscopically, or fluorometrically; B represents a non-reactive organic bridging unit linking the moiety Q to a reactive functional group f, said bridging unit being of sufficient length or size to prevent any possible interaction of Q that might alter its spectroscopic properties even upon derivatization; and f is a reactive group which can react with a compound to form covalently bonded derivatives.

Abstract: A novel reactor for reacting and subsequently analyzing sub-picomole quantities of a sample organic molecule. The reactor includes a continuous capillary connected between two valves that control fluid flow in the capillary. One part of the capillary forms a reaction chamber where the sample may be immobilized for subsequent reaction with reagents supplied through the valves. Another part of the capillary passes through or terminates in the detector portion of an analyzer such as an electrophoresis apparatus, liquid chromatographic apparatus or mass spectrometer. The apparatus may form a peptide or protein sequencer for carrying out the Edman degradation reaction and analyzing the reaction product produced by the reaction.

Abstract: Disclosed herein are surface activated, organic polymers useful for biopolymer synthesis. Most preferably, aminated biaxially oriented polypropylene is used for the synthesis of oligonucleotides thereto, and these devices are most preferably utilized for genetic analysis of patient samples.

Abstract: A method for sequencing proteins on a polytetrafluoroethylene support is described. The support is preferably porous. The sample to be sequenced may be transferred directly, e.g., by blotting, to the support. Covalent binding of the sample to the support is not required.

Abstract: A method for correlating a peptide fragment mass spectrum with amino acid sequences derived from a database is provided. A peptide is analyzed by a tandem mass spectrometer to yield a peptide fragment mass spectrum. A protein sequence database or a nucleotide sequence database is used to predict one or more fragment spectra for comparison with the experimentally-derived fragment spectrum. In one embodiment, sub-sequences of the sequences found on the database which define a peptide having a mass substantially equal to the mass of the peptide analyzed by the tandem mass spectrometer are identified as candidate sequences. For each candidate sequence, a plurality of fragments of the sequence are identified and the masses and m/z ratios of the fragments are predicted and used to form a predicted mass spectrum.

Abstract: To easily detect an amino acid derivative with high sensitivity in carrying out amino acid sequence analysis of a protein from an amino terminal, 2-anilino-5-thiazolinone amino acid derivative is reacted with a volatile primary amine represented by the general formula X-NH.sub.2, wherein X denotes a hydrocarbon containing a halogen to form a phenylthiocarbamyl amino acid derivative, and the phenylthiocarbamyl amino acid derivative is detected by using a gas chromatograph provided with an ECD. Accordingly, amino acid sequencing may be performed with high sensitivity without the need to use harmful radioisotope labeled or fluorescent amino compounds.

Abstract: The present invention provides methods and reagents for sequencing amino acids. One embodiment of the method for determining the terminal amino acid of a polypeptide comprises the steps of (a) attaching (either covalently or non-covalently) the polypeptide to a solid support, (b) reacting the polypeptide with a compound described below, under conditions and for a time sufficient for coupling to occur between the terminal amino acid of the polypeptide and the compound, thereby yielding a polypeptide with a derivatized terminal amino acid, (c) washing the solid support to remove unbound material, (d) cleaving the derivatized terminal amino acid from the polypeptide with a cleaving agent, (e) ionizing the cleaved derivatized terminal amino acid, and (f) determining the molecular weight of the derivatized terminal amino acid, such that the terminal amino acid is determined.Within one embodiment, the compound is p-isothiocyanato phenethyl trimethylammonium and counterion salts thereof.

Abstract: The invention provides a method for determining an amino acid sequence motif for a phosphorylation site of a protein kinase. In the method of the invention, a protein kinase is contacted with an oriented degenerate peptide library, peptides within the library which are substrates for the kinase are converted to phosphopeptides and the phosphopeptides are separated from non-phosphorylated peptides. The isolated phosphopeptides are sequenced and an amino acid sequence motif for the phosphorylation site is determined based upon the relative abundance of different amino acids residues at each degenerate position. The invention also provides peptide substrates for protein kinase A, cell cycle control kinases, src family kinases, the EGF receptor and p92.sup.c-fps/fes based upon amino acid sequence motifs for the phosphorylation sites of these kinases.

Abstract: An N-terminal peptide sequencing method is disclosed in which a thiocarbamoyl compound is reacted with N-terminal amino acid of the sample to form a derivative of said amino acid which is then cleaved.

Abstract: A method for scanning nucleotide or DNA sequence data banks to identify genetic regions or genes coding for biologically interacting proteins.

Abstract: A protein or peptide is treated by a vapor containing an organic acid represented by the following general formula CF.sub.3 --(CF.sub.2)n--COOH (n is zero or more integer). The resulting reaction mixture is processed by a mass spectrometer to obtain a mass spectrum to measure a mass of respective chemical species contained in the reaction mixture. Alternatively, the reaction mixture is processed by an amino acid analyzer to determine an amino acid sequence of the protein or peptide from the carboxy-terminal. According to another method, the protein or peptide is treated by an anhydride of the organic acid represented by the following general formula CF.sub.3 --(CF.sub.2)n--COOH (n is zero or more integer). The resulting reaction mixture is processed by a mass spectrometer to obtain a mass spectrum to measure a mass of respective chemical species contained in the reaction mixture to determine an amino acid sequence of the protein or peptide from the carboxy-terminal.

Abstract: A first reactant is immobilized i.e. in a porous matrix (50), adjacent a sample electrode (46) within a reaction chamber. Energizing of the electrode (46) electrophoretically attracts a mobile second reactant and/or electrolytically induces appropriate reaction conditions to enhance reaction of the first and second reactants. Polarity reversals between the sample electrode (46) and remote electrodes (38), (42), (44) cause unreacted second reactant and/or by-products to migrate away from the immobilized first reactant. The techniques are useful for sequential chemical reactions such as sequencing or construction of proteins, polysaccharides and nucleic acids where cyclical additions and removals of reactants are required. The techniques are amenable to automated micro and nano scale construction and operation and allow direct electrophoretic (38) interfacing with chromatographic, HPCE and mass spectrophotometric equipment.

Abstract: The instant invention provides a library of bio-oligomers of defined size and known composition, in which the library contains all of the possible sequences of the bio-oligomers, and a method of synthesis thereof. The bio-oligomers of the library may be peptides, nucleic acids, or a combination of the foregoing. The instant invention also provides methods to identify bio-oligomers from a library that demonstrate desired characteristics such as binding, bioactivity and catalytic activity. Thus the instant invention provides a unique and powerful method to identify a useful bio-oligomer sequences from a library more quickly than current state-of-the-art technology allows. Effector molecules for use in treatment or diagnosis of disease are also provided.

Abstract: In many separation techniques, such as field flow fractionation, liquid chromatography and electro-phoresis, chemical species form bands that migrate at different velocities. If the data-digitization rate and excitation intensity are both set to be optimal for the fastest migrating band, to compensate for different band velocities, both the data-digitization rate and the excitation intensity are decreased as a function of time by a factor equal to the migration time of the fastest migrating band to the separation time.

Abstract: Processes are provided for pretreating body fluid compositions and subsequently analyzing the pretreated body fluid compositions for analytes of interest. Processes for pretreating the compositions include providing size exclusion gel having a molecular weight fractionation range or a molecular weight exclusion such that the size exclusion gel is capable of excluding or fractionating the analytes of interest, and then causing the composition to contact the size exclusion gel in order to separate the analytes from low molecular weight composition components which interfere with the separation and analysis of the analytes of interest. Processes for analyzing pretreated compositions include electrophoretic methods such as capillary zone electrophoresis which involve the separation and detection of analytes of interest.

Abstract: A method for quantitatively measuring an acid in a polar organic solvent involves adding a sterically hindered base, such as diisopropylentylamine, triethylamine or N-methylmorpholine to the solvent and measuring the conductance of the mixture. The method has particular relevance in monitoring the progress of preactivation, coupling and deprotection reactions in solid phase peptide synthesis (SPPS).

Abstract: The present invention relates to a method for peptide C-terminal fragment sequence analysis, in which the fragment collection is carried out on an allylamine group-derivatized polymer membrane or on allylamine group-derivatized glass fiber filter paper; the collected C-terminal fragment is immobilized thereon using a water-soluble carbodiimide etc.; and the obtained immobilized product is subjected directly to amino acid sequence analysis. The present invention also relates to an apparatus for collecting a peptide fragment. According to the method of the present invention, peptides which are rich in hydrophobic groups in their C-terminus and are therefore difficult to trap with polyvalent ion carriers, currently used in the gas-phase sequencer, can be completely analyzed up to their C-terminus. Also, amino acid sequence analysis can be made even when the amount of C-terminal fragments is very small.

Abstract: The present invention relates to methods for determining the amino acid composition, and more preferably the sequence, of a peptide using mass spectrometric techniques. The method is particularly useful for sequencing peptides isolated from natural sources or from libraries of peptides that have been prepared synthetically, and for peptides that are not amenable to Edman degradation sequencing. In one embodiment, the method for determining the amino acid composition or sequence of a peptide comprises determining the difference of the mass of the peptide from the mass of a deuterium-hydrogen exchanged peptide, and from this difference determining the number of exchangeable (labile) hydrogen atoms (protons). Candidate peptides having amino acid compositions or sequences that do not contain the observed number of exchangeable protons are eliminated.

Abstract: A method is provided for C-terminal sequencing of a protein or peptide. An important feature of the method is the formation of an oxazolone moiety at the C-terminus of a protein or peptide by treatment with acetic anhydride under basic conditions followed by conversion of the oxazolone to a thiohydantoin moiety by treatment with thiocyanate under acidic conditions. Yields of thiohydantoin are further enhanced by delivering thiocyanate as the conjugate acid of a sterically hindered alkylammnonium cation.

Abstract: Method and device for the synthesis of macromolecules, including peptides, polynucleotides or oligosaccharides. The method and device involves a reactor which contains a solid support on which a component of the product to be synthesized is fixed. The reactor comprises filters at its ends and is mounted movably for rectilinear reciprocating movement in a vessel which contains a reactive liquid. A temperature probe is provided to detect the temperature variations of the reactive liquid due to passage into and out of the reactor. The progress of coupling and deprotection reactions which occur in the synthesis of peptides, polynucleotides and oligosaccharides is monitored.

Abstract: A cleavage apparatus for obtaining a free peptide after assembly of a peptide on a solid support by solid-phase peptide synthesis equipped with a cleavage cocktail transport line installed between the column for cleavage containing a peptidyl resin and a cleavage cocktail supply bottle, through which a cleavage cocktail is supplied from the bottle to the column and recovered into the cleavage cocktail supply bottle from the column. This apparatus can provide easy, safe, rapid and reliable cleavage in a large scale peptide synthesis.

Abstract: Improved methods for the GC/MS analysis of sulfhydryl amino acids and methylmalonic acids in samples of body fluids are provided. Additionally a method for combined assay of sulfhydryl amino acids particularly total homocysteine and combined creatine/creatinine levels in samples of body fluids provided. The information provided by the methods described is useful in the detection of the presence of cobalamin or folate deficiencies in individuals and in distinguishing between the two deficiencies.

Abstract: A computer-assisted method for identifying protein sequences that fold into a known three-dimensional structure. The inventive method attacks the inverse protein folding problem by finding target sequences that are most compatible with profiles representing the structural environments of the residues in known three-dimensional protein structures. The method starts with a known three-dimensional protein structure and determines three key features of each residue's environment within the structure: (1) the total area of the residue's side-chain that is buried by other protein atoms, inaccessible to solvent; (2) the fraction of the side-chain area that is covered by polar atoms (O, N) or water, and (3) the local secondary structure. Based on these parameters, each residue position is categorized into an environment class.

Abstract: A carboxy terminal protein sequencing process is disclosed which utilizes phosphoroisothiocyanatidate for the derivatization step.

Abstract: An improved method for the N-terminal sequential degradation of proteins and peptides is disclosed. The protein or peptide to be sequenced is reacted with a compound effective to impart a tertiary amine functionality to the thiazolinone derivative of a cleaved terminal amino acid of the peptide.

Abstract: A novel method for the sequential degradation from the N-terminus of small samples of proteins or peptides. The released amino acids may be detected by mass spectrometry.

Abstract: A method for detecting elevated levels of the middle isoform of .alpha.-2 haptoglobin, .alpha.-2FS haptoglobin in bodily fluids of subjects with Alzheimer's disease and schizophrenia as compared with the .alpha.-2FS haptoglobin level in normal subjects. Fibrinogen fragments corresponding to proteins 127 and 128 are also found in elevated levels in such subjects. Elevated levels of specific haptoglobin proteins serve as a diagnostic marker for Alzheimer's disease and Schizophrenia.

Abstract: The subject invention concerns novel materials and methods for the detection, treatment, and prevention of human osteoarthritis. Specifically, the cleavage site where aggrecanase cleaves aggrecan has been identified. Identification of this site, as well as the nature of the enzyme, facilitates specific treatments which block or diminish the activity of the enzyme. A further aspect of the invention concerns methods for detecting evidence of osteoarthritis.

Abstract: A novel reactor for reacting and subsequently analyzing sub-picomole quantities of a sample organic molecule. The reactor includes a continuous capillary connected between two valves that control fluid flow in the capillary. One part of the capillary forms a reaction chamber where the sample may be immobilized for subsequent reaction with reagents supplied through the valves. Another part of the capillary passes through or terminates in the detector portion of an analyzer such as an electrophoresis apparatus, liquid chromatographic apparatus or mass spectrometer. The apparatus may form a peptide or protein sequencer for carrying out the Edman degradation reaction and analyzing the reaction product produced by the reaction.

Abstract: The present invention is directed to the implementation of a multi-port rotary valve in an automated chemistry processing instrument which reduces chemical reagent cross contamination and simplifies system design and control. One or more multi-port rotary valves are used in conjunction with isolation valves which are each dedicated for an associated reagent in the system. According to one embodiment of the present invention, the instrument utilizes a multi-port valve which defines several fluid branches each associated with a reagent. The valve has a common inlet and common outlet which are selectively brought into fluid communication with the branches in a controlled sequence. At each branch, there is a two-way three-port isolation valve which controls introduction of an associated reagent into the branch. When a branch is selected by the rotary valve, the reagent introduced into the branch is delivered out of the common outlet by the flow from the common inlet.

Abstract: A method for determining three-dimensional structural information of a protein involves producing the protein in a form substantially labeled with .sup.13 C or .sup.15 N or both and subjecting the protein to nuclear magnetic resonance spectroscopic analysis. The isotopically labeled protein is produced by a method which involves producing a substantially labeled microbial protein hydrolysate, subjecting the protein hydrolyzate to cation exchange chromatography to produce a partially purified labeled amino acid mixture, subjecting the partially purified labeled amino acid mixture to anion exchange chromatography to produce a purified labeled amino acid mixture and supplementing the purified labeled amino acid mixture with isotopically labeled cysteine.

Abstract: The instant invention provides a library of bio-oligomers of defined size and known composition, in which the library contains all of the possible sequences of the bio-oligomers, and a method of synthesis thereof. The bio-oligomers of the library may be peptides, nucleic acids, or a combination of the foregoing. The instant invention also provides methods to identify bio-oligomers from a library that demonstrate desired characteristics such as binding, bioactivity and catalytic activity. Thus the instant invention provides a unique and powerful method to identify a useful bio-oligomer sequences from a library more quickly than current state-of-the-art technology allows. Effector molecules for use in treatment or diagnosis of disease are also provided.

Abstract: A method for screening and distinguishing between cobalamin deficiency and folic acid deficiency by relating elevated levels of cystathionine to cobalamin or folic acid deficiency and relating elevated levels of 2-methylcitric acid to cobalamin deficiency but not folic acid deficiency. The methods can be used alone or in combination with other methods for detecting and distinguishing between cobalamin deficiency and folic acid deficiency.

Abstract: The present invention is directed to an automated peptide synthesizer equipped with recovery lines including an acyl component recovery line for recovering unreacted acyl components to allow the reuse of the acyl components, and a solvent recovery line for recovering and distilling a reaction solvent and passing it through a purification column to allow the reuse of the reaction solvent. Both recovery lines are installed on the flow pathway from one or more reaction chambers or columns to a waste liquid reservoir of the automated peptide synthesizer. By recovering expensive acyl components and solvents, running cost in peptide synthesis can be reduced to mitigate economic burden, and the amount of waste leading to environmental destruction can be reduced.

Abstract: A method of rational drug design includes simulating polypeptides in a way that predicts the most probable secondary and/or tertiary structures of a polypeptide, e.g., an oligopeptide, without any presumptions as to the conformation of the underlying primary or secondary structure. The method involves computer simulation of the polypeptide, and more particularly simulating a real-size primary structure in an aqueous environment, shrinking the size of the polypeptide isobarically and isothermally, and expanding the simulated polypeptide to its real size in selected time periods. A useful set of tools, termed Balaji plots, energy conformational maps, and probability maps, assist in identifying those portions of the predicted peptide structure that are most flexible or most rigid. The rational design of novel compounds, useful as drugs, e.g., bioactive peptidomimetic compounds, and constrained analogs thereof, is thus made possible using the simulation methods and tools of the described invention.