TY - JOUR
T1 - The production of high affinity monoclonal antibodies to human growth hormone
AU - Stuart, Margaret C.
AU - Walichnowski, Christine M.
AU - Anne Underwood, P.
AU - Hussain, Safinaaz
AU - Harman, Deirdre F.
AU - Rathjen, Deborah A.
AU - Von Sturmer, Shona R.
PY - 1983/6/24
Y1 - 1983/6/24
N2 - 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.
AB - 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.
KW - human growth hormone
KW - monoclonal antibody
KW - radioimmunoassay
UR - http://www.scopus.com/inward/record.url?scp=0020631131&partnerID=8YFLogxK
U2 - 10.1016/0022-1759(83)90005-4
DO - 10.1016/0022-1759(83)90005-4
M3 - Article
C2 - 6854038
AN - SCOPUS:0020631131
SN - 0022-1759
VL - 61
SP - 33
EP - 42
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
IS - 1
ER -