Resonance Scattering Spectral Study of IgA Immune Complex Particles and Its Analytical Application
LIANG Ai-hui1,WANG Na2,HOU Ming1
1. Department of Material and Chemical Engineering, Guilin University of Technology, Key Laboratory of New Processing Technology for Nonferrous Metals and Materials, Ministry of Education, Guilin 541004, China 2. Department of Resource and Environmental Science, Guangxi Normal University, Guilin 541004, China
Abstract：Based on the resonance scattering (RS) effect of immune complex particles, a new resonance scattering spectral method for the determination of trace IgA in the human blood serum was developed. It was based on that goat anti-human IgA was combined with the antigen of IgA. It is known that antibody has C-terminal and N-terminal and the N-terminal is binding site of antigen, and it could combine antigen. Because the stereo structure anastomoses and the charge are opposite between goat anti-human IgA and IgA, they could attract and combine each other. The attraction and combination have high idiosyncrasy and they are done by Val der Waals force, hydrophobic force, Coulomb attracting force and hydrogen bond binding force, and aggregate to form immune complex particles that exhibit three resonance scattering peaks at 340，390，420，450，470 and 520 nm respectively, in the pH 6.2 Na2HPO4-citrate buffer solutions and in the presence of polyethylene glycol (PEG) 4000. The laser scattering results indicate that the average diameter of the immune complex particles is about 1 100 nm. The influences of pH, PEG type and its concentration, goat anti-human immunoglobulin A concentration, incubation temperature and incubation time, foreign substance such as glucose, human serum albumin (HSA), urea and L-tyrosine were examined in details. Under the chosen conditions of pH 6.2 Na2HPO4-citrate buffer solutions-60 mg·mL-1 polyethylene glycol(PEG) 4000-0.35 mL anti-human immunoglobulin，the resonance scattering intensity at 340 nm ΔI340 nm and the resonance scattering intensity at 470 nm ΔI470 nm all are both proportional to the concentration of IgA in the range of 0.133-4.67 μg·mL-1. Its regress equation was ΔI340 nm=18.61 cIgA+3.19, ΔI470 nm=18.57cIgA+ 6.51，with a detection limit of 0.068 and 0.072 μg·mL-1,respectively. The assay has been applied to the analysis of IgA in human serum, which was also determined by the immunoturbidimetric method, with satisfactory results. The slope, intercept and correlation coefficient of the linear regress analysis were 1.064, -0.213 and 0.929 9, respectively.
[1] YAO Chun-yan, FU Wei-ling(姚春艳，俯伟灵). Chinese New Medicine(中国新医药)，2004，3(3)：55. [2] LIU De-sheng(刘德生). J. Wuhan Medicine College(武汉医学院学报)，1985，3：206. [3] Hocini Hakim, Iscaki Sylvio, Bouvet Jean-Pierre, et al. Journal of Immunological Methods，2000, 235: 53. [4] WANG Shi-ping(王世平). Chinese Journal of Sport Medicine(中国运动医学杂志)，2001，20(4)：392. [5] Wu X, Sun S N, Yang J H, et al. Spectrochimica Acta Part A, 2005, 62：896. [6] Zhiliang Jiang, Sumei Zhou, Aihui Liang, et al. Environ. Sci. Technol., 2006, 40: 4286. [7] BAI Yan, LI Wei-jia, WU Ya-qin，et al(白 燕，李维嘉，吴雅琴, 等). Spectroscopy and Spectral Analysis(光谱学与光谱分析), 2006，26(2)：313. [8] ZHANG Biao-ming, JIANG Zhi-liang, ZHAI Hao-ying, et al(章表明，蒋治良，翟好英，等). Spectroscopy and Spectral Analysis(光谱学与光谱分析)，2005, 25(7): 1092. [9] Rutao Liu, Jianghe Yang, Changxia Sun, et al. Microchimica Acta, 2004, 147(1-2): 105. [10] Wu X, Li L, Yang J H, et al. Microchim Acta, 2003, 141: 165. [11] Li Y X, Wu Y Q, Chen J L, et al. Anal. Bioanal. Chem., 2003, 377: 675. [12] Liu S P, Luo H Q, Li N B, et al. Anal. Chem., 2001, 73: 3907. [13] L P Wu, Y F Li, C Z Huang, et al. Anal. Chem., 2006, 78: 5570. [14] LIU Shao-pu, JIANG Zhi-liang, KONG Ling(刘绍璞，蒋治良，孔 玲). Science in China, Ser/ B Chemistry(中国科学，B辑)，2002，32(6)：614. [15] JIANG Zhi-liang, CHEN Yuan-yuan, LIANG Ai-hui, et al(蒋治良，陈媛媛, 梁爱惠, 等). Science in China, Ser. B Chemistry(中国科学，B辑)，2006, 36(5): 419. [16] Chen H Q, Xu F G, Hong S, et al. Spectrochimica Acta Part, A, 2006, 65(2)：428. [17] Zhiliang Jiang, Shuangjiao Sun, Aihui Liang, et al. Clin. Chem., 2006, 52: 1389. [18] DENG Jun-yao, SUN Shuang-jiao, JIANG Zhi-liang, et al(邓俊耀, 孙双姣, 蒋治良, 等). Spectroscopy and Spectral Analysis(光谱学与光谱分析), 2006，26(8)：1487.
