Preparation and characteristics of Ca(2+)-dependent monoclonal antibodies to recoverin.

Thirty-four primary hybridoma clones were prepared which expressed monoclonal antibodies to the Ca(2+)-binding protein recoverin. Among the resulting monoclonal antibodies, two Ca(2+)-dependent clones (mAb3 and mAb19) recognizing recoverin were detected by solid-phase immunoenzyme assay. In the presence of Ca(2+), antibodies of the mAb3 and mAb19 clones bound to recoverin several times better than in the absence of Ca(2+). The mAb3 and mAb19 antibodies recognized epitopes located inside the sequences Pro61-Met91 and Pro57-Tyr64 of the recoverin molecule, respectively. The possible mechanism of the Ca(2+)-dependent recognition of recoverin by the prepared monoclonal antibodies is discussed.