Isolation of antihemorrhagic factors in opossum (Didelphis virginiana) serum using a monoclonal antibody immunoadsorbent
| Autor: | John C. Perez, Shyi Yi Huang, Shu-Fang Tarng |
|---|---|
| Rok vydání: | 1986 |
| Předmět: |
Didelphis
medicine.drug_class Antihemorrhagic Enzyme-Linked Immunosorbent Assay Toxicology Monoclonal antibody Chromatography Affinity Sepharose Mice Affinity chromatography medicine Animals Polyacrylamide gel electrophoresis Mice Inbred BALB C Chromatography biology Chemistry Isoelectric focusing Antibodies Monoclonal Blood Proteins Opossums biology.organism_classification Molecular biology Molecular Weight Sephadex Electrophoresis Polyacrylamide Gel Isoelectric Focusing Immunosorbents |
| Zdroj: | Toxicon : official journal of the International Society on Toxinology. 24(6) |
| ISSN: | 0041-0101 |
| Popis: | The antihemorrhagic factor in opossum (Didelphis virginiana) serum isolated by Sephadex G-200 gel filtration and DEAE A-50 ion exchange chromatography was used as antigen to immunize BALB/c mice. Hybrid cell lines secreting monoclonal antibodies against antihemorrhagic factor were produced by fusion of Sp2/0 myeloma cells with spleen cells of the immunized mice. The ascites fluid was produced in BALB/c mice. The monoclonal antibody in the ascites fluid was partially purified by DEAE A-50 ion exchange and coupled to CNBr-activated 4B Sepharose to be used as an immunoadsorbent in affinity chromatography for isolation of antihemorrhagic factor. The neutralization capacity of the convetionally isolated antihemorrhagic factor was 14.6 times and the affinity isolated antihemorrhagic factor was 16.8 times that of crude opossum serum. Both antihemorrhagic factors were homogeneous, with one fast migrating band in the area of albumin shown by polyacrylamide gel electrophoresis. However, the antihemorrhagic factor showed one heavy band and one faint band in SDS-polyacrylamide electrophoresis, as well as in isoelectric focusing. The molecular weight of the heavy band was estimated to be 65,000 with a value of pI 4.8 and the faint band was 57,000 with a value of pI 4.1. |
| Databáze: | OpenAIRE |
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