An alternate method utilizing small quantities of ligand for affinity purification of monospecific antibodies.

Abstract:

:An alternate method was designed to couple a limited quantity of protein to an affinity support when a conventional technique was unsuccessful. This was achieved through the introduction of a small number of sulfhydryl groups to the ligand by reaction with 2-iminothiolane which resulted in a limited number of reactive sites on the protein. Amino groups on an AH-Sepharose 4B matrix were linked to sulfhydryl groups on the ligand using the heterobifunctional agent m-maleimidobenzoyl sulfosuccinimide ester (sulfo-MBS). This method was employed to prepare an affinity support using a cytosolic protein that activates glyceraldehyde-3-phosphate dehydrogenase as a ligand. Monospecific antibody purified from the affinity column recognized only this protein on a Western blot of a cytosolic extract of kidney epithelial cells.

journal_name

J Immunol Methods

authors

Aithal HN,Knigge KM,Kartha S,Czyzewski EA,Toback FG

doi

10.1016/0022-1759(88)90034-8

subject

Has Abstract

pub_date

1988-08-09 00:00:00

pages

63-9

issue

1

eissn

0022-1759

issn

1872-7905

pii

0022-1759(88)90034-8

journal_volume

112

pub_type

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