The production of high affinity monoclonal antibodies to human growth hormone

Margaret C. Stuart*, Christine M. Walichnowski, P. Anne Underwood, Safinaaz Hussain, Deirdre F. Harman, Deborah A. Rathjen, Shona R. Von Sturmer

*Corresponding author for this work

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Abstract

The primary aim of this work was to produce specific monoclonal antibodies to human growth hormone (hGH) for use in a diagnostic RIA of hGH levels in serum. Three different schedules were used for immunization of BALB/c mice and the splenocytes from each mouse were fused with myeloma cells Sp 2/0 Ag 14. Each fusion resulted in the production of hundreds of hybridomas secreting hGH-directed antibodies. Six antibodies have been fully characterized and have been grouped into pairs which recognize 3 different epitopes on the hGH molecule. One pair exhibits no cross reaction with the structurally related placental hormone, human placental lactogen (hPL), a second pair has low cross reaction with hPL (1.6-3%) and a third pair reacts equally well with hGH and hPL indicating binding to a common epitope in the 2 molecules. The highest affinity antibody, 74 6, which has an affinity constant of 4.4 × 1010 1/mol and 3% cross-reactivity with hPL, has been used to establish a RIA for serum hGH measurements. Evidence is provided that hGH levels measured in this assay correlate well with those obtained in a conventional rabbit antiserum assay.

Original languageEnglish
Pages (from-to)33-42
Number of pages10
JournalJournal of Immunological Methods
Volume61
Issue number1
DOIs
Publication statusPublished - 24 Jun 1983

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Keywords

  • human growth hormone
  • monoclonal antibody
  • radioimmunoassay

Cite this

Stuart, M. C., Walichnowski, C. M., Anne Underwood, P., Hussain, S., Harman, D. F., Rathjen, D. A., & Von Sturmer, S. R. (1983). The production of high affinity monoclonal antibodies to human growth hormone. Journal of Immunological Methods, 61(1), 33-42. https://doi.org/10.1016/0022-1759(83)90005-4