Monoclonal antibodies to human growth hormone can distinguish between pituitary and genetically engineered forms |
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Authors: | R Aston L Cooper A Holder J Ivanyi M Preece |
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Institution: | 1. Department of Experimental Immunobiology, Wellcome Research Laboratories, Langley Court, Beckenham, Kent, U.K.;3. Department of Growth and Development, Institute of Child Health, University of London, 30 Guildford Street, London WC1N 1EH, U.K. |
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Abstract: | Monoclonal antibodies (MABs) prepared against human pituitary growth hormone (hGH) have been compared for their binding to pituitary-derived and genetically engineered methionyl growth hormone (met-hGH). The antibodies bind to four non-overlapping epitopes of which two are completely shared with human choronic somatomammotropin (hCS). The determinant defined by MAB NA27 was expressed on met-hGH to a lesser degree than on hGH of pituitary origin. However, another antibody, QA68, which binds to a determinant closely related to NA27, failed to discriminate between hGH and met-hGH. A further two MABs (EB1 and NA71) were similarly ineffective in distinguishing between the two forms of the hormone. The determinant recognized by antibody EB2 was equally represented on hGH and met-hGH when assessed by a liquid-phase radioimmunoassay: however, measurement of the binding in a solid-phase assay resulted in a two-four-fold lower binding to met-hGH. Bioactivity assessed by both an in vitro cell proliferation assay and an in vivo cartilage sulphation bioassay failed to distinguish between the two hormones. It is therefore concluded that the NH2-terminal methionine on bacterially derived growth hormone results in altered antigenicity of the hormone without any measurable effect on bioactivity. |
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Keywords: | hGH human growth hormone met-hGH bacterially derived methionyl hGH (rDNA-hGH) hCS human chorionic somatomammotropin PBS phosphate-buffered saline FCS foetal calf serum HS horse serum RIA radioimmunoassay KDa kilodaltons |
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