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US 5,350,674 AGranted
Intrinsic factor - horse peroxidase conjugates and a method for increasing the stability thereof
AssigneeBecton, Dickinson and Company
InventorsMichael T. Boenisch, Erica A. Sullivan
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Abstract
This invention presents a non-isotopic competitive assay for Vitamin B12 (B12), utilizing intrinsic factor (IF) labelled with horse radish peroxidase (HRP), by coupling via heterobifunctional cross-linking agents. In addition, a method for stabilizing the resultant conjugates by pretreatment with N-ethylmaleimide is disclosed.
Claims7
A method for producing a stabilized HRP-SATA-SMPB-IF conjugate comprising:
- (i) admixing intrinsic factor (IF) with N-ethylmaleimide (NEM) to form an NEM/IF product;
- (ii) reacting said NEM/IF product with succinimidyl-4- (p-maleimidophenyl) butyrate (SMPB) to form a SMPB-IF product;
- (iii) reacting horseradish peroxidase (HRP)with N-succinylimidyl-5-acetylthioacetate (SATA) to form a SATA-HRP product, and subsequently deacetylating said product to form a deacetylated SATA-HRP product;
- (iv) admixing said SMPB-IF product with said deacetylated SATA-HRP product to form a HRP-SATA-SMPB-IF conjugate;
- and
- (v) subsequently concentrating said conjugate.
3 more elements
Dependencies are inferred from the claim text.