Preparation and Characteristics of Ca²⁺-Dependent Monoclonal Antibodies to Recoverin

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N. K. Tikhomirova¹, M. A. Goncharskaya², and I. I. Senin^1*

¹Laboratory of Visual Reception, Department of Cell Signal Systems, Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119992 Moscow, Russia; fax: (7-095) 939-0978; E-mail: senin@genebee.msu.su

²Laboratory of Immunochemistry, Institute of Carcinogenesis, Blokhin Cancer Research Center, Russian Academy of Medical Sciences, Kashirskoe Shosse 24, 115478 Moscow, Russia

^* To whom correspondence should be addressed.

Received December 1, 2003; Revision received March 24, 2004

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Thirty-four primary hybridoma clones were prepared which expressed monoclonal antibodies to the Ca²⁺-binding protein recoverin. Among the resulting monoclonal antibodies, two Ca²⁺-dependent clones (mAb3 and mAb19) recognizing recoverin were detected by solid-phase immunoenzyme assay. In the presence of Ca²⁺, antibodies of the mAb3 and mAb19 clones bound to recoverin several times better than in the absence of Ca²⁺. 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 Ca²⁺-dependent recognition of recoverin by the prepared monoclonal antibodies is discussed.

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KEY WORDS: recoverin, Ca²⁺-binding proteins, phototransduction, monoclonal antibodies, recoverin epitopes

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