Radioimmunoassay of glycated serum protein using monoclonal antibody to glucitollysine and coomassie-brilliant-blue-coated polystyrene beads.
A radioimmunoassay for glycated serum protein (GSP) was developed using monoclonal antibody to glucitollysine and polystyrene beads coated with Coomassie-Brilliant-Blue (CBB) as adsorbent for serum protein. The monoclonal antibody was raised by immunizing BALB/c mice with reduced glycated LDL and fusing their spleen cells with mouse myeloma cells. CBB-coated polystyrene beads were introduced to absorb a constant amount of serum protein. The protein adsorbed on the CBB-coated beads was reduced by NaHB4, and after treatment with radiolabeled antibody, the radioactivity of each bead was counted with an automatic gamma-counter. The standard glycated protein used was reduced glycated human serum albumin, in which 8 of 59 lysine residues were glycated. The intra- and interassay coefficients of variation of GSP were 4.8-6.5% and 1.6-6.0%, respectively. The GSP level of diabetic patients was significantly higher than that of normal controls (1.97 +/- 1.23 vs. 0.47 +/- 0.21 nmol/mg-protein; mean +/- SD, p less than 0.001). The GSP levels of patients with insulin-dependent and non-insulin-dependent diabetes mellitus were 3.03 +/- 1.05 and 1.51 +/- 1.00 nmol/mg-protein, respectively. A good correlation was found between the levels of GSP and hemoglobin A1c (HbA1c) (r = 0.85, p less than 0.001). In patients admitted to the hospital for diabetes education and glycemic control, the GSP level decreased 43 +/- 12% with the decrease in the fasting plasma glucose level (39 +/- 13%) and the mean daily plasma glucose level (MPG, 47 +/- 15%) in a four week period after admission, whereas the HbA1c level decreased only 13 +/- 6% during this period.(ABSTRACT TRUNCATED AT 250 WORDS)
Yamamoto
Y
Y
Department of Geriatric Medicine, Osaka University Medical School, Japan.
Tahara
Y
Y
Cha
T
T
Noma
Y
Y
Fukuda
M
M
Yamato
E
E
Yoneda
H
H
Hashimoto
F
F
Ohboshi
C
C
Hirota
M
M
D000818:Animals; D000911:Antibodies, Monoclonal; D001798:Blood Proteins / Q000032:analysis*; D003922:Diabetes Mellitus, Type 1 / Q000097:blood*; D003924:Diabetes Mellitus, Type 2 / Q000097:blood*; D006442:Glycated Hemoglobin / Q000032:analysis; D017127:Glycation End Products, Advanced; D006023:Glycoproteins; D006031:Glycosylation; D006801:Humans; D007202:Indicators and Reagents; D008239:Lysine / Q000031:analogs & derivatives* / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D011137:Polystyrenes; D011863:Radioimmunoassay / Q000379:methods; D012016:Reference Values; D012394:Rosaniline Dyes; D012709:Serum Albumin / Q000032:analysis; D000093422:Glycated Serum Proteins; D000093423:Glycated Serum Albumin
D000911:Antibodies, Monoclonal; D001798:Blood Proteins; D006442:Glycated Hemoglobin; D017127:Glycation End Products, Advanced; D006023:Glycoproteins; D007202:Indicators and Reagents; D008239:Lysine; D011137:Polystyrenes; D012394:Rosaniline Dyes; D012709:Serum Albumin; D000093422:Glycated Serum Proteins; D000093423:Glycated Serum Albumin; C034247:glucitollysine; C004692:coomassie Brilliant Blue