Purification and Characterization of Transforming growth factor - beta1 from Human Platelets / 대한면역학회지
Korean Journal of Immunology
;
: 1-8, 1998.
Article
in Korean
| WPRIM
| ID: wpr-90172
ABSTRACT
Transforming growth factor-j31 (TGF-p1) has potential for therapeutic use in common clinical conditions for which there are no adequate pharmacological agents. However, in vivo studies using TGF-p1 were hindered by high price of this cytokine. As a first step towards large scale purification of TGF-p1, it was purified in a small scale (10 unit platelets) from human platelets by four purification steps:
platelet extraction, gel filtration, cation exchange chromatography, and reversed phase high performance liquid chromatography (HPLC). A single protein band with a molecular weight of 25 Kd corresponding to purchased TGF-p1 (R8D Systems) was confirmed by silver staining after SDS-polyacrylamide gel electrophoresis (SDS-PAGE) of eluant from reversed phase HPLC. Recovery (%) of each step was about 50-60%, resulting in the final recovery of 20% based on the detection by a sandwich ELISA. Approximately, 3.7 p,g of purified TGF-p1 was obtained from 18 pg of platelet extracts. This result was confirmed by receptor (TGF-j31 type II) ELISA and bioassay using a mink lung epithelial'cell line (MV1LU). Further, in vitro characterization study showed that purified TGF-p1 inhibits G1/S transition of LPS-activated murine spleen B cells and increases surface IgA expression by the same cell population, which are typical activities of TGF-p1 in B cell differentiation. Taken together, the results from the present study reveals that purified TGF-p1 is fully biologically active and our purification methodology could be usbful to obtain a large scale of recombinant TGF-p1 in the future.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Spleen
/
Biological Assay
/
Blood Platelets
/
Immunoglobulin A
/
Enzyme-Linked Immunosorbent Assay
/
B-Lymphocytes
/
Transforming Growth Factors
/
Cell Cycle
/
Cell Differentiation
/
Chromatography
Limits:
Humans
Language:
Korean
Journal:
Korean Journal of Immunology
Year:
1998
Type:
Article
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