A competitive enzyme linked immunosorbent assay for recombinant Bm86 produced in Pichia pastoris

Authors: Reytor E.¹; Hernández L.²; Santana H.²; Blanco R.¹; Barceló M.T.¹; Rodríguez I.³; Boué O.²

Source: Biotechnology Techniques, Volume 12, Number 12, December 1998 , pp. 919-923(5)

Publisher: Springer

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Abstract:

A competitive enzyme linked immonosorbent assay (ELISA) was developed for quantification of recombinant Bm86 protein (rBm86), antigen of the vaccine GavacTM against the cattle tick Boophilus microplus. Monoclonal antibody (Mab) SSBm1 showed identical recognition to folded and denatured antigen and was used in the competition assay. This ELISA was sensitive enough to detect 60 ng/ml of rBm86. Within-assay coefficient of variation was 5.7 to 6.2 % and between-assay variation was 7.7 to 11.6 %. The level of expression of recombinant Bm86 in Pichia pastoris reached about 2.7 g per liter of culture after 80 hour of fermentation. © Rapid Science Ltd. 1998

Keywords: ELISA; Pichia pastoris; cattle tick; vaccine (Gavac); recombinant protein

Language: English

Document Type: Regular paper

Affiliations: 1: Sancti Spiritus, Ave 31 % 158 y 190, Cubanacán Havana Cuba 2: Havana, Ave 31 % 158 y 190, Cubanacán Havana Cuba 3: Center for Genetic Engineering and Biotechnology, Ave 31%158 y 190, Cubanacán Havana Cuba

Publication date: 1998-12-01