Elimination of ingredients effect to improve the detection of anti HIV-1 p24 antibody in human serum using SPR apparatus.
Highly sensitive detection of proteins in serum becomes difficult in some cases during surface plasmon resonance (SPR) measurements, because some ingredients in the serum hugely enhance non-specific reactions on the sensing chip of SPR. It is well recognized that the antibody against core protein p24 of HIV in serum is one of the most important proteins in the accurate diagnosis of infection with HIV. In this study, we could attain the accurate detection of anti p24 antibody in human serum by eliminating the serious effects of the ingredients in serum on the measurement of SPR by employing these procedures: 1) blocking the gold surface of the sensing chip with human serum and 2) heating the serum sample at 56 degrees C for 30 min. Without these treatments, the signal of SPR was considerably suppressed on the measurements of the anti p24 antibody which contained human serum, making the accurate detection difficult. However, with introducing the above two treatments, the sensing of anti p24 antibody in human serum was improved, while a small non-specific reaction was still observed. By removing the non-specific reaction caused by the ingredients in the serum, we could accurately measure the antibody for p24 in human serum sample over the range from 1 to 20 micrograms/ml.
Hiroshima Prefectural University, School of Biosciences, Shobara, Hiroshima 727-0023, Japan.
Emi
E
Hifumi
Naruhiko
N
Kubota
Yoshiyuki
Y
Niimi
Kosuke
K
Shimizu
Naoyoshi
N
Egashira
Taizo
T
Uda
D000818:Animals; D000911:Antibodies, Monoclonal / Q000097:blood; D002021:Buffers; D002138:Calibration; D004867:Equipment Design; D015483:HIV Antibodies / Q000097:blood*; D016655:HIV Core Protein p24 / Q000276:immunology*; D006801:Humans; D051379:Mice; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D020349:Surface Plasmon Resonance / Q000592:standards*
D000911:Antibodies, Monoclonal; D002021:Buffers; D015483:HIV Antibodies; D016655:HIV Core Protein p24