Situ process for production of conjugates

• US 5,324,650 A
• Filed: 09/17/1992
• Issued: 06/28/1994
• Est. Priority Date: 03/20/1990
• Status: Expired due to Fees

• Current Assignee: Dade Behring, Inc. (Siemens AG)
• Original Assignee: E. I. du Pont de Nemours and Company (Corteva, Inc.)
• Inventors: Tseng, Susan Y., Obzansky, David M.
• Primary Examiner(s): Robinson, Douglas W.
• Assistant Examiner(s): Weber, Jon P.
• Application Number:
  US07/946,247
• Time in Patent Office: 649 Days
• Field of Search: 424/85.8, 424/94.3, 435/7.1, 435/7.6, 435/188, 435/964
• US Class Current: 435/188
• CPC Class Codes: C12N 9/96 Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
• CPC Class Codes: G01N 33/531 Production of immunochemical test materials
• CPC Class Codes: G01N 33/532 Production of labelled immunochemicals
• CPC Class Codes: Y10S 435/964 including enzyme-ligand conjugate production, e.g. reducing rate of nonproductive linkage

Abstract

In situ process for production of conjugates using a deblocking agent which allows conjugation to occur and two compositions wherein one composition is functionalized with a group capable of reacting efficiently with free thiols, and a second composition containing blocked thiols.

Claims

1. A process for conjugating two compounds which comprises mixing simultaneously (i) a first compound functionalized with a maleimide, (ii) a second compound containing blocked thiols, and (iii) a deblocking reagent wherein said deblocking reagent is hydroxylamine to generate free thiol groups wherein said simultaneous mixing is such that deblocking of the thiol and conjugation to the maleimide functionalized compound occurs substantially simultaneously and further wherein the first and second compounds can be the same or different and are selected from the group consisting of proteins, peptides, nucleotides and lipids derivatives with at least one amine group.

2. A process according to claim 1 wherein the first compound is functionalized with a heterobifunctional cross linking agent selected from the group consisting of SMCC, SMBP, and MBs and the second compound is functionalized with a thiolating reagent selected from the group consisting of SATP, SATA, and SAMSA.