Abstract: The present invention relates to a novel class of bioconjugates derived from phenylboronic acid complexes, and the method of making and using such bioconjugate complexes. The complexes are in the form of the following general formulas. ##STR1## wherein group Q is selected from either O, S, NH, N-alkyl and N-aryl, wherein alkyl denotes a hydrocarbon moiety, wherein aryl is selected from either an aromatic ring, a substituted aromatic ring and fused aromatic rings, wherein group R is preferably selected from either O, NH, CH.sub.2, alkyl and aryl, wherein alkyl and aryl are as were previously defined,wherein group X is selected from either H, CH.sub.3 and C.sub.6 H.sub.5, wherein group Y is selected from either O, NH, CH.sub.

Abstract: The present invention relates to a novel class of phenylboronic acid complexing reagents useful for the preparation of bioconjugates, and the method of making and using such reagents. In the present invention, in the place of prior art Avidin-Biotin and Digoxigenin-anti-Digoxigenin systems, phenylboronic acid complexing reagents are utilized in conjunction with phenylboronic acid reagents (many of which are known in the prior art) to facilitate chemical conjugation without the use of intermediary biological macromolecules. Reagents suitable for the modification of a bioactive species for the purpose of incorporating a phenylboronic acid complexing moiety for subsequent conjugation to a different bioactive species having pendant phenylboronic acid moleties are of General Formula I or General Formula II. ##STR1## wherein group X is selected from either H, OH, NH.sub.2, NHCH.sub.3, NHOH and NHOCH.sub.

Abstract: The present invention relates to a novel class of phenylboronic acid bioconjugate complexes derived from aminosalicylic acid, and the method of making and using such bioconjugate complexes. The bioconjugate complexes are in the form of the following general formulas; ##STR1## wherein group X is selected from the group consisting of O, NH, N-alkyl, NC.sub.6 H.sub.5, N-aryl, NCH.sub.2 -aryl, NCH.sub.2 CH.sub.2 OH, NCOCH.sub.2 CH.sub.2 OH, NOH, NO-alkyl and NOCH.sub.2 -aryl, wherein alkyl denotes a hydrocarbon moiety of from 1 to 4 carbons in length and which may be linear or branched, wherein aryl is selected from the group consisting of an aromatic ring, a substituted aromatic ring and a fused aromatic ring,wherein group Y is selected from the group consisting of O, S, NH, N-alkyl, N-aryl and NCH.sub.

Abstract: The present invention comprises a reaction cell for a sequencer and the like which includes a sample carrier assembly having a flexible, resilient spacer with a hole therein for retaining the sample on a membrane, a pair of alumina cells which bracket the spacer, one or more pads or spacers to retain the assembly in a tight fitting relation and a cap to secure the assembly together. The reagents are input and withdrawn from opposite edges of the sample membrane. The present invention has a minimal dead volume as a result of the spacer arrangement. It also permits benchtop sample loading due to the separation of the assembly into a carrier and base housing. The fluid lines are self-sealing to the reaction cell.

Abstract: A protein microsequencing method for use in conjunction with the thiobenzoylation 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 thiobenzoylating reagent. The derivatized polypeptide is subjected to cleavage by acid which forms a 4-substituted 2-phenyl-5(4H) thiazolone. The thiazolone is acylated to form a 5-acyloxy-2-phenylthiazole and subjected to detection by both gas chromatography and chemical ionization mass spectroscopy.

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: The invention relates to the functionalization of particulate bonded phase chromatographic supports prepared by silanization of silica gel or controlled pore glass and containing pendant primary alkyl amine groups. Functionalization results from the activation of the amines by reaction with N,N'-carbonyldiimidazole (CDI), or a related azolide, in anhydrous organic solvent, followed by derivatization of the support. Derivatization results from reaction of the activated support with a functionalizing reagent consisting of a primary or secondary, alkyl or aryl amine in organic solvent, or from an aqueous solution of the amine or its salt. A urea linkage results through which the functionalizing reagent is covalently attached to the support. Derivatization can result from the addition of an excess of a single reagent, or as a consequence of the sequential addition of two or more functionalizing reagents.

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 solid phase oxime compound having the general formula: ##STR1## where X is particulate silica gel or controlled pore glass; n is 3 or 4, n' is 1-6; R' is an electron withdrawing group; and Y and Y' are selected from OH, OCOR", OSO.sub.2 R" and OCO.sub.2 R", where R" is a chromophore, fluorophore or electrophore, is disclosed. Also disclosed is the method for making compounds of this type and the method for using these compounds to detect and quantify analytes having primary or secondary amines or thiol functionalities.

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.