Analytical Sciences

Elimination of Ingredients Effect to Improve the Detection of Anti HIV-1 p24 Antibody in Human Serum Using SPR Apparatus

Emi HIFUMI,* Naruhiko KUBOTA,* Yoshiyuki NIIMI,* Kosuke SHIMIZU,* Naoyoshi EGASHIRA,* and Taizo UDA*^,** 

*Hiroshima Prefectural University, School of Biosciences, Shobara, Hiroshima 727-0023, Japan
**CREST of JST (Japan Science and Technology Corporation)

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°C for 30 min. Without these treatments, the signal of SPR was considerably suppressed on the measurement 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 ug/ml.

J-STAGE: View this article in J-STAGE