Modification of T-cell antigenic properties of tetanus toxoid by SDS-PAGE separation. Implications for T-cell blotting
Research output: Contribution to journal › Journal article › Research › peer-review
Using Tetanus Toxoid (TT) as a model antigen the T-cell Blotting method was evaluated. Peripheral blood mononuclear cell (PBMC) cultures were stimulated by blotted nitrocellulose-bound TT or soluble TT. SDS-Poly-Acrylamide-Gel-Electrophoresis separated TT only induced proliferation in 20% of the PBMC cultures whereas proliferation was induced in 79% of the same cultures offered similar treated TT (except for the PAGE separation). When T-cell blotting was performed with TT separated in a SDS-agarose matrix, proliferation was induced in 80% of donors responding to soluble TT. The results show that SDS-PAGE alters the ability of TT to induce T-cell proliferation, possibly due to unpolymerized acrylamide binding to proteins during SDS-PAGE. The use of SDS-PAGE T-cell blotting in the screening for T-cell antigens must therefore be reconsidered. We suggest the use of SDS-Agarose Gel Electrophoresis as an alternative when doing T-cell blots.
Original language | English |
---|---|
Journal | Journal of Immunoassay and Immunochemistry |
Volume | 18 |
Issue number | 2 |
Pages (from-to) | 129-48 |
Number of pages | 19 |
ISSN | 1532-1819 |
Publication status | Published - 1997 |
Bibliographical note
Keywords: Acrylamide; Acrylamides; Collodion; Electrophoresis, Agar Gel; Electrophoresis, Polyacrylamide Gel; Epitopes, T-Lymphocyte; Humans; Immunoblotting; Lymphocyte Activation; Solubility; T-Lymphocytes; Tetanus Toxoid
ID: 6766257