An alternate method utilizing small quantities of ligand for affinity purification of monospecific antibodies
| Autor: | K. M. Knigge, Aithal Hn, Toback Fg, Czyzewski Ea, Sreedharan Kartha |
|---|---|
| Rok vydání: | 1988 |
| Předmět: |
Immunology
Dehydrogenase Monospecific antibody Chromatography Affinity Matrix (chemical analysis) Western blot Affinity chromatography Antibody Specificity medicine Animals Humans Immunology and Allergy biology medicine.diagnostic_test Chemistry Antibodies Monoclonal Glyceraldehyde-3-Phosphate Dehydrogenases Ligand (biochemistry) Immunohistochemistry Cytosol Cross-Linking Reagents Biochemistry Immunoglobulin G biology.protein Female Binding Sites Antibody Rabbits Antibody |
| Zdroj: | Journal of Immunological Methods. 112:63-69 |
| ISSN: | 0022-1759 |
| DOI: | 10.1016/0022-1759(88)90034-8 |
| Popis: | 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 actives 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. |
| Databáze: | OpenAIRE |
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