Abstract: A computer based system and method determines and displays possible chemical structures for converting two naturally aggregated but chemically separated polypeptide chains into a single polypeptide chain which will fold into a three dimensional structure very similar to the original structure made of the two polypeptide chains. A data base contains a large number of amino acid sequences for which the three dimensional structure is known. After plausible sites have been selected, this data base is examined to find which amino acid sequences (linkers) can bridge the gap between the plausible sites to create a plausible one-polypeptide structure. The testing of each possible linker proceeds in three steps. First, the span (a scalar quantity) of the candidate is compared to the span of the gap. If the span is close enough, step two is done which involves aligning the first peptides of the candidate with the initial peptide of the gap.

Abstract: A method for detecting amino acid derivatives in which 2-anilino-5-thiazolinone derivatives of amino acids are reacted with radioactive iodine siotope-labeled amino compounds or with fluorescent amino compounds to form phenylthiocarbamyl amino acid derivatives. The phenylthiocarbamyl amino acid derivatives are detected with a high level of sensitivity by a radioactive detector or a fluorescence spectrophotometer.

Abstract: A sequencing method which exploits the thioacylation degradation of polypeptides and proteins is disclosed. The process involves reaction of the N-terminal amino acid of a polypeptide with an excess of a thioacylating reagent. After sufficient time to insure quantitative coupling and removal of excess reagent, the N-thioacyl polypeptide is subjected to cleavage by acid which affords a 2-substituted-5(4H)-thiazolinone of the N-terminal amino acid. Subsequent addition of a large excess of an aliphatic primary or secondary alcohol, either directly to the cleavage acid or after its removal, yields the corresponding N-thioacyl amino acid ester, a stable compound suitable for chromatographic separation and subsequent detection by contemporary methods of high pressure liquid chromatography.

Abstract: A protein micro sequencing method for use in conjunction with the thiocylation degradation of polypeptides and proteins is disclosed. The process involves reaction of the N-terminal amino acid of a polypeptide with an excess of a thioacylating reagent. After sufficient time to insure quantitative coupling and removal of excess reagent, the derivatized polypeptide is subjected to cleavage by acid which affords a 2-substituted-5(4H)-thiazolinone. After removal of excess acid, the thiazolinone is reacted with a small excess of a fluorescent or enhanced ultraviolet absorbance reagent having a reactive carboxylic acid chloride, sulfonic acid chloride, chloroformate, isocyanate or anhydride functionality, in the presence of a tertiary amine catalyst, to yield the corresponding 5-O-acyl-2-(substituted)thiazole derivative, detectible by enhanced ultravoilet absorobance or fluorescence emission at extremely low concentration thereby providing a method of sequencing very small amounts of protein.

Abstract: According to one aspect of the invention operationally macrocyclic molecular structures are employed to input and/or to read out information, electrical potential, optical characteristics, energy levels, state information, etc. with respect to another material, such as a neurological or biological material, nucleic acid or other materials. In one example disclosed in the above mentioned application macrocyclic molecules were adsorbed on a silver substrate; in the present invention the operationally macrocyclic molecular structure is adsorbed onto other substrates, such as nerve cells, muscles, nucleic acid (RNA and DNA), etc.

Abstract: A computer-based method evaluates the structure of a protein to thereby identify sites in the protein molecule at which the natural amino acid residues can be replaced with cysteine residues in order to permit the formation of a potentially stabilizing disulfide bond.

Abstract: An apparatus and process are used to cyclicly degrade a peptide to be sequenced, arriving at a set of amino acid residues for each cycle. The amount of each amino acid residue is quantitatively measured in each set, then a background level is fit to each cycle to obtain a background fit. A measure of dispersion is then calculated for the background fit, and the measured amounts of amino acid residues in each cycle are normalized relative to the background fit. The largest normalized background-corrected residue amount in each cycle then provides a sequence assignment that can be used for further correction steps if desired.

Abstract: A method of assaying primary amines or cyanide is disclosed. The process is carried out by reacting a substituted or unsubstituted naphthalene-2,3-dicarboxaldehyde or o-phthalaldehyde, reactive with primary amines and cyanide, with cyanide ions and at least one primary amine to form an adduct amenable to fluorometric or electrochemical assaying techniques. The adduct is then fluorometrically or electrochemically assayed.Also disclosed are 1-cyano-benz[f]isoindole adducts.

Abstract: A plurality of analytes are analyzed by liquid chromatography in a liquid biological sample containing many components which can interfere with analysis by liquid chromatography. The sample, containing the analytes of interest, are passed to a first passage of a dialyzer which removes molecules in the sample which interfere with analysis. A diluent is passed through a second passage of the dialyzer in which the two liquids are separated from each other by a semipermeable membrane having holes large enough to permit the analytes of interest to pass into the diluent but holes small enough to prevent passage of sample components which interfere with analysis. The analyte-containing diluent is passed to a retaining element for temporarily retaining the analytes of interest by adsorption and/or partition. The analytes of interest are removed from the retaining element by a second diluent which is then passed through a chromatography column with a detector and the analytes are analyzed chromatographically.

Abstract: A peptide is sequenced from its carboxyl terminal by coupling with an isothiocyanate having a chemical formula R(.sub.x)Si(NCS).sub.y where x=0-3 and y=1-4. R may be alkyl or aryl. The coupling reaction is preferably conducted at a temperature range of 50.degree.-90.degree. C. for a period to approximately sixty (60) minutes or less. A weak acid anhydride, preferably a carboxylic acid anhydride, or a weak acid chloride, preferably a carboxylic acid chloride, may be mixed with the isothiocyanate. An organic base catalyst selected from a group consisting of tertiary amines including aromatic amines such as pyridine may also be utilized.

Abstract: A probe and method for using the same for supporting a sample in an ion source of a mass spectrometer comprises a target formed by a receptacle for a liquid sample in which the sample passes by diffusion to the high vacuum side of a semipermeable membrane and/or in which the sample is held as a droplet by surface tension. In order to replenish the droplet surface, the supply can be arranged to overflow the receptacle.

Abstract: The invention relates to a new chiral reagent of the formula ##STR1## wherein X is halogen, an azide group or a succinimidyl group and wherein R is an alkyl group or a trifluormethyl group. The invention further relates to a method for the derivatization, determination and preparation resolution of primary and secondary amino-containing compounds. The method comprises the steps of derivatizing the amino function by the reagent to form diastereomeric carbamates, which are determined by e.g. fluorimetry or absorptiometry after separation by liquid chromatography.

Abstract: A solid phase N-hydroxysuccinimide reagent having the general formula: ##STR1## where X is particulate silica gel or controlled pore glass; n=3-7; Y is either (CH.sub.2).sub.n' or (CH.sub.2).sub.n' --NH--CO--CH.sub.2 wherein n'=2-6; Z may be in the form of OCOR, OSO.sub.2 R and OCO.sub.2 R, and Z' may be either Z or OH, wherein R is a chromophore, fluorophore or electrophore capable of detection in chromatographic separation technology is disclosed. Also disclosed is the method for making compounds of this type and the method for using these compounds to identify and quantitate analytes having primary or secondary amine or thiol functionalities.

Abstract: A method for monitoring a solid-phase peptide synthesis for free amino ends of unreacted peptide chains, the monitoring occurring at the end of each addition of a blocked amino acid to the peptide chains which are anchored to the solid phase. The method includes reacting the solid phase with a monitoring agent that forms a covalent bond with unblocked amino groups. The solid phase is then washed to remove the unreacted monitoring agent. A cleaving reagent is then used to selectively remove the covalently bound monitoring reagent from the ends of the unblocked peptide chains, while leaving the blocked chains intact. The amount of monitoring agent thus removed is then quantitatively measured to determine what proportion of the initial peptide chains failed to react with the blocked amino acid.Preferred monitoring agents for use in this process include trityl (triphenylmethyl) -based groups, and particularly preferred agents are trityl, monomethoxytrityl, dimethoxytrityl, and trimethoxytrityl chlorides.

Abstract: A mixture of fluorescently-excitable amino acid derivatives suitable for use in quantitative amino acid analysis is provided in which primary amino acids are present as derivatives of a first reagent and secondary amino acids are present as derivatives of a second different reagent. The process for preparing the mixture comprises the steps of derivatization of the primary amino acids of an amino acid sample with ortho-phthalaldehyde (OPA) and of derivatization of the secondary amino acids of the sample with fluorenylmethylchloroformate (FMOC) in the presence of acetonitrile. The mixture can be obtained automatically and is highly suitable for fast analysis by reversed-phase liquid chromatography.

Abstract: Polypeptides are disclosed which inhibit the binding of the C5a chemotaxin to polymorphonuclear leukocytes by cleaving a six amino acid peptide from the carboxy-terminus of the C5a chemotaxin. The polypeptides can be isolated from virulent strains of group A streptococci, s. pyogenes, by enzymatic or detergent extraction.

Abstract: A chemiluminescence method for assaying compounds containing primary amino groups using 1-cyano-2-substituted benz(f)- or naphth(f)-isoindole fluorescers is disclosed. The 1-cyano-2-substituted benz(f)- or naphth(f)-isoindole (CBI or CNI) is formed by reacting a compound containing a primary amino group with naphthalene-2-3-dicarboxaldehyde or anthracene-2,3-dicarboxaldehyde in the presence of cyanide ion. The reaction product of hydrogen peroxide with an oxalate ester is then combined with the CBI or CNI to form an analyte which exhibits chemiluminescence. A detector is used to measure the chemiluminescence emission from the chemiluminescence derivatized analytes. Compounds containing primary amino groups which may be assayed according to the present invention include primary amines, amino acids, peptides and catecholamines. The method can also be adapted to the analysis of trace amounts of cyanide ion and hydrogen peroxide.

Abstract: A method is provided for quantitatively monitoring the deprotection and coupling reactions employed in the solid phase synthesis of peptides. The method entails synthesizing a peptide on a support matrix that has a first marker associated therewith. The amino acids employed in the peptide synthesis procedure have a second marker attached thereto, which can be the blocking group used for the amino acid. After the coupling or deprotection step a portion of the support matrix is processed to release first and second identifers from the first and second markers, respectively. The completeness of the coupling or deprotection step can be determined by comparing the relative amounts of the detected first and second identifiers. Novel compositions of matter are used in or produced during this method, including support matrixes having pyrolyzable markers attached thereto.

Abstract: An apparatus for peptide synthesis in which a plurality of supply valves are arranged in series to define a line for supplying successive reagents to a reaction vessel in which a peptide is to be synthesized. A corresponding plurality of containers contain respective reagents and each, by actuation of its corresponding valve, is connectable through downstream ones of the series of valves to supply its respective reagent to the reaction vessel. For cleaning or reacting with peptide starting material therein further valving permits draining of the reaction vessel to waste and, between applications of successive reagents to the reaction vessel, cleaning of the line of valves by flushing with a cleaning reagent sent from the upstream end of the series of valves through the line to waste.

Abstract: Apparatus for carrying out a multiplicity of different sequential chemical syntheses on solid-supports simultaneously comprises a multiplicity of complementary support plates each formed with a reaction chamber opening through the plate and containing the solid-support together with at least one by-pass channel through the plate. The by-pass channel or channels and reaction chamber are disposed equi-angularly and at the same radial distance about an axis of the plate. The plates are rotatably supported in compression in face to face contact between end plates complementary to the plates and each formed with a blank position and passages for at least one reactant stream. Means is provided for independently rotating each plate whereby each reaction chamber of each plate may independently be isolated or positioned in the or a selected reactant stream.

Abstract: A method of forming analyzable adducts in a mixture of compounds by contacting the mixture with a boron reagent having the formula of either ##STR1## where each X and Y is, independently, an alkyl group of 12 or fewer carbons or an aryl group of 6-20 carbons; or BZ.sub.3, where each Z is, independently, an alkyl group of 12 or fewer carbons, or an aryl group of 6-20 carbons.

Abstract: A method for fluorometric analysis of catecholamines characterized in that, after the addition of a catecholamine into a solution of 1,2-diphenylethylenediamine containing an oxidizing reagent at a pH of 4 to 10, the mixture is allowed to stand for more than 1 minute at a temperature of higher than 15.degree. C. to make cateocholamine fluorescent, is disclosed.

Abstract: A computer based system and method determines, and displays possible chemical structures for converting two naturally aggregated but chemically separated polypeptide chains into a single polypeptide chain which will fold into a three dimensional structure very similar to the original structure made of the two polypeptide chains. A data base contains a large number of amino acid sequences for which the three dimensional structure is known. After plausible sites have been selected, this data base is examined to find which amino acid sequences (linkers) can bridge the gap between the plausible sites to create a plausible one-polypeptide structure. The testing of each possible linker proceeds in three steps. First, the span (a scaler quantity) of the candidate is compared to the span of the gap. If the span is close enough, step two is done which involves aligning the first peptides of the candidate with the initial peptide of the gap.

Abstract: An improved apparatus and method for the sequential performance of chemical processes on a sample of chemical material wherein the sample is embedded in a solid matrix of fluid permeable material located within a reaction chamber and is sequentially subjected to a plurality of fluids passed through the chamber in a pressurized stream, causing chemical interaction between the sample and the fluids.

Abstract: Methods are provided for determining the presence or absence of ragged ends in polymeric proteins or protein products, by means of fast atom bombardment and high mass mass spectrometry (preferably high field magnet mass spectrometry). The methods may be modified for locating N- or C-terminal peptide in a polymeric protein or protein product and for assigning disulphide bridges in polymeric protein or protein products. The methods are applicable to other biopolymers such as nucleotides and carbohydrates.

Abstract: A system maintained under a constant reference pressure for the automated synthesis of peptides includes a reaction vessel that has a single port for both injection and withdrawal of the various fluids used in the peptide synthesis sequence, a plurality of reservoirs for holding the amino acids used in the synthesis of the peptide chains and a plurality of reservoirs for holding the solvents and reagents used to promote the synthesis of the peptide chains. The system also includes a volume displacement pump for removing a controlled volume of gas from the reaction vessel as the first part of each injection step to reduce the pressure within the vessel. This is followed by connection of a selected reservoir to the reaction vessel and flow from the reservoir to the vessel resulting from the pressure differential between the vessel and the reservoir. As soon as the pressure in the reaction vessel has returned to reference level, the transfer is complete.

Abstract: A method of amino acid analysis by reacting an amino acid-containing sample with o-phthalaldehyde and an N-protective group-substituted cysteine such as N-acetyl-L-cysteine under an alkaline condition to yield a fluorescent compound and measuring the intensity of fluorescence or absorbance of the resulting compound. The method is sensitive for quantitative and qualitative analysis of a secondary amino acid such as proline when it is beforehand subjected to an oxidation.

Abstract: A rapid method for determining the isoelectric point for an amphoteric molecule of interest is provided which utilizes the pH-dependent binding affinity of the molecule for an ion-exchange material. The empirically derived pI values are within a range of 0.2 pH units or less of the reported values in the literature and may be more precisely determined by employing a narrower pH gradient as part of the procedure. The unique methodology allows isoelectric point determinations to be accomplished within one hour's time, avoids carrier ampholyte interaction or artifact formation, and allows the determination to be performed at any desired temperature with decreased risk of denaturation.

Abstract: A method of sequencing polypeptides is disclosed utilizing liquid-solid affinity chromotography. The method utilizes a reagent reactive at one position with the amino moiety of a terminal amino acid, and reactive at a second position with boronic acid. The reagent is coupled to the terminal amino acid. A scavenger molecule having a sulfhydryl group and an amine group reacts with the excess reagent and the excess scavenger and scavenger-reagent complex are removed on an immobilized organomercurial column. The terminal amino acid is cleaved from the polypeptide, and coupled to an immobilized boronic acid column. The amino acid is removed from the column and identified and the remainder polypeptide is recycled.

Abstract: A method of analyzing molecular layers of amino-functional group coupling agents on inorganic substrates is disclosed. A saturated solution of 1-dimethylaminonaphthalene-5-sulfonyl chloride is added to a sample to be tested, and sufficient time is allowed, with or without heating, for the reaction of the 1-dimethylaminonaphthalene-5-sulfonyl chloride with any amino functional group coupling agent which might be present. The color intensity of the reaction product is compared under ultraviolet light in an otherwise substantially dark room with the color intensity of standards representative of known thicknesses of coupling agent to determine the existence and thickness of the coupling agent on the particles being analyzed.

Abstract: Conversion flask for use is an apparatus for the sequential performance of chemical processes on a sample of chemical material. The flask includes at least three capillary tubes and a large bore tube extending into the interior thereof for the introduction and withdrawal of various fluids. The capillary tubes are constructed to introduce a plurality of fine bubbles into a liquid within the flask to agitate the liquid and accelerate drying, to impinge chemicals on the walls of the flask, and to produce a spray onto the interior walls of the flask in proximity to its upper end to wash down the interior walls of the flask.

Abstract: An improved method for the sequential performance of chemical processes on a sample of chemical material wherein the sample is embedded in a solid matrix of fluid permeable material located within a reaction chamber and is sequentially subjected to a plurality of fluids passed through the chamber in a pressurized stream, causing chemical interaction between the sample and the fluids.

Abstract: A microprocessor controlled, general purpose gene synthesizer for programmably synthesizing selected nucleotide sequences. Depending upon a programmed chemistry, metered volumes of selected bases and reagent/solvents are sequentially metered into the bottom of a reaction cell containing solid-supported nucleotides, where desired linkages are achieved in an agitated suspension. Self-refilling syringe metering and electrically initiated, pneumatically operated valving insures economical, non-contaminated synthesis. Preprogrammed, selectable chemistries and operator programmed sequences are operatively coupled under microprocessor control to ensure a general purpose synthesizer.

Abstract: A protein sequencing method utilizing a composition of matter which comprises a D-C-B-A reagent wherein A is a moiety which can react with and bind to a terminal amino acid of a protein and can result in removal of the terminal amino amino acid, B is a moiety which provides steric separation between C and A, C is a nucleophilic moiety which can be detected, and D is a moiety which protects the C moiety from degradation or other modification, and is labile in acidic media and stable in neutral or basic media. The protein sequencing method and reagent can be used in micro sequencing apparatus.

Abstract: A diagnostic process is disclosed for detecting very early or developed stages of malignancy of tissue beneath a predetermined portion of the skin of a human patient. A sample of body secretion is obtained from the outer surface of the skin and is tested for presence of markedly decreased free, naturally occurring amino acids. The test has approximately the same order of sensitivity as a standard ninhydrin test. The diagnostic process of the invention is particularly adapted for mass screening of human females for early stages of breast cancer.

Abstract: A temperature control apparatus for a high temperature reactor in an analytical instrument of the type in which a development reagent is added to a flowing stream as part of the detection process. The temperature control apparatus utilizes solid state heat transfer means to transfer heat between a thermoconductive block to which is mounted the reactor and an air exchange heat sink which obtains heat from or disposes heat into ambient air as required. The control apparatus regulates the temperature of the reactor at a very rapid rate in the high temperature range of approximately 90.degree. C. to 150.degree. C. without the use of a separately contained heat exchange medium. The ability of the apparatus to cool the reactor rapidly eliminates the requirement to keep reactor contents at elevated pressures to prevent vaporization of the stream being analyzed.

Abstract: A process for quantitatively assaying factor Xa from a medium by reacting the medium with tripeptide derivatives having the formula ##STR1## wherein R.sup.5 is a chromogenic substituted amino group capable of being split off by enzymatic hydrolysis to form a colored or fluorescent product H--R.sup.5. The quantity of split product H--R.sup.5 released by the enzymatic action on the tripeptide derivative is determined photometrically, spectrophotometrically, fluorescence-spectrophotometrically, or electrochemically. The quantity of released split product H--R.sup.5 per time unit is proportional to the quantity of enzyme present in the starting material.

Abstract: An improved method for analyzing amino acids by silylation with silylating agents of the formulae: ##STR1## wherein R is lower alkyl, Y is hydrogen, lower alkyl or perfluoroloweralkyl and Y.sub.1 is hydrogen or lower alkyl.

Abstract: Antineoplastons are termed a group of plasma, tissue and urinary peptides and amino acid derivatives capable of modulating abnormal tissue growth, such as neoplastic disease. Antineoplastons, when administered to persons with neoplastic disease, have been shown to be effective against several forms of cancer and tumors. A procedure is provided herein for the determination of antineoplaston levels in physiological tissues or fluids, especially plasma and urine. The procedure involves purification of antineoplastons by high performance liquid chromatography on silica gel followed by resolution of antineoplastons by high performance liquid chromatography on sulfonated polystyrene. A quantitative determination of antineoplaston tissue or fluid levels provides valuable data to aid clinical diagnosis of cancer. In addition, the quantitative determination also provides a means for monitoring the efficacy of antineoplastic therapy.

Abstract: A method for detecting fluorescent materials in liquid chromatography. Fluorescent materials are excited by chemical reaction with chemical reagents to yield light (chemiluminescence) and detected at high sensitivity. The detection apparatus is composed of a mixer for mixing a solution containing separated fluorescent materials and solutions of chemical reagents to afford chemiluminescence, a flow cell and a light receptive detection means for detecting the light thus produced.