A simple method for the preparation and purification of C1 complement cleaved beta 2-microglobulin from human serum.
Research output: Contribution to journal › Journal article › Research › peer-review
A simple method is described for the preparation of proteolytically processed forms of beta 2-microglobulin suitable for structural and biological studies. PEG 6000 was added to the serum of healthy individuals to precipitate the C1 complement complex from C1 esterase inhibitor (C1-inh). After dissolving the precipitate containing the C1 complement in Tris-HCl buffer, pH 7.6, efficient conversion of added beta 2-microglobulin to desLys58 beta 2-microglobulin was observed. Addition of a specific carboxypeptidase B inhibitor (Plummers inhibitor) could partly prevent the deletion of Lys-58 from cleaved beta 2-microglobulin, whereby Lys58-cleaved beta 2-microglobulin was obtained. The proteolytically processed forms were subsequently purified by G-75 Sephadex gel filtration followed by chromatofocusing. A yield of 10-40% of proteolytically processed beta 2-microglobulin was obtained. Only one component was seen by SDS-PAGE stained with Coomassie Brilliant Blue.
Original language | English |
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Journal | Journal of Immunological Methods |
Volume | 205 |
Issue number | 1 |
Pages (from-to) | 29-33 |
Number of pages | 4 |
ISSN | 0022-1759 |
Publication status | Published - 1997 |
Bibliographical note
Keywords: Chromatography, Gel; Complement C1; Enzyme-Linked Immunosorbent Assay; Humans; beta 2-Microglobulin
ID: 8746501