Monoclonal antibodies reveal circulating growth hormone of high molecular weight not detectable by conventional assays.
A radioimmunoassay based on a monoclonal antibody. Mc-ab 1, which was raised against growth hormone but cross-reacted with human placental lactogen yielded higher GH immunoreactivity levels in serum than one based on a polyclonal antiserum. This discrepancy was noted in subjects with normal GH secretion as well as in patients with GH insufficiency. To characterize this GH immunoreactivity detected by Mc-ab 1, affinity purification and molecular sieve chromatography of serum were performed. High molecular weight proteins with GH immunoreactivity were found with both techniques. These proteins were associated with carbohydrates. Affinity cross-linking showed specific binding of radiolabelled GH to high molecular weight proteins in the serum. After fractionation of serum, the GH immunoreactivity became detectable by the polyclonal antiserum assay as well as by an immunoradiometric assay. GH immunoreactive material with an approximate mass of 80 kD was subjected to isoelectric focusing. When GH immunoreactive fractions at pH 5 were re-chromatographed, GH immunoreactivity was recovered in the elution volume corresponding to monomeric GH. Our results show that sera from normal subjects as well as from patients with deficient GH secretion contain notable amounts of high molecular weight GH which is undetectable by antibodies generally used for GH measurements, but which can be revealed after fractionation of serum.
Luthman
M
M
Department of Endocrinology, Karolinska Hospital, Stockholm, Sweden.
Jónsdóttir
I
I
Skoog
B
B
Wivall
I L
IL
Roos
P
P
Werner
S
S
D000172:Acromegaly / Q000378:metabolism; D000328:Adult; D000368:Aged; D000911:Antibodies, Monoclonal; D002846:Chromatography, Affinity; D002940:Circadian Rhythm; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D013006:Growth Hormone / Q000097:blood*; D006801:Humans; D007018:Hypopituitarism / Q000378:metabolism; D007525:Isoelectric Focusing; D008297:Male; D008875:Middle Aged; D010902:Pituitary Gland / Q000378:metabolism; D015175:Prolactinoma / Q000378:metabolism; D011863:Radioimmunoassay
D000911:Antibodies, Monoclonal; D013006:Growth Hormone