| 宋晶晶,罗 萍,王 娇,周水梅,沈长新.免疫磁珠技术分离唾液A/B血型抗原并用于吸附血清A/B抗体[J].现代生物医学进展英文版,2018,(14):2617-2622. |
| 免疫磁珠技术分离唾液A/B血型抗原并用于吸附血清A/B抗体 |
| Separation of Blood Group Antigen A/B by Immunomagnetic Separation and Utilization of the Adsorption Blood Group A/B Antibodies |
| Received:October 13, 2017 Revised:November 08, 2017 |
| DOI：10.13241/j.cnki.pmb.2018.14.004 |
| 中文关键词: 磁性免疫分离 血型抗原A 血型抗原B 免疫吸附 |
| 英文关键词: Immunomagnetic separation Blood group A antigen Blood group B antigen Immunoadsorption |
| 基金项目:湖北省卫生计生委采供血专项基金项目（WJ2015CB005） |
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| 中文摘要: |
| 摘要 目的：应用免疫磁珠分离技术获得具有良好抗原性的A/B血型抗原，并探究其作为ABO血型抗体吸附剂去除A/B抗体的可行性。方法：将含有血型物质的唾液进行预处理，再与包被了抗体的磁珠混合，分离出纯度较高的A/B抗原，运用酶联免疫及凝集抑制试验验证所得抗原的抗原性及是否存在交叉反应。用未纯化A/B抗原和纯化A/B抗原包被磁珠，对含有抗A/B IgM、IgG的血清进行抗体吸附，用纯化A/B抗原对100份来自O型血孕妇的临床血清样本进行抗体吸附，分别评价其吸附效果。结果：纯化抗原与对应抗体反应后，其吸光度显著高于对照组(A抗原与A抗体0.85±0.12 vs. 0.27±0.03，P<0.01；B抗原与B抗体0.86±0.09 vs. 0.24±0.06，P<0.01)，与其它类型抗体反应后的吸光度值与对照组比较差异无统计学意义(P>0.05)。进行红细胞凝集抑制试验时，纯化抗原可显著抑制相应抗体与红细胞的凝集反应，对其它类型抗体与红细胞的凝集没有抑制作用。血清抗体吸附实验表明纯化抗原的吸附效率比未纯化抗原的高(97.00 % vs. 88.00 %，P<0.001)。临床样本抗体吸附实验显示，纯化A抗原对抗A IgM/IgG的吸附效率分别为96.88 %、98.44 %；纯化B抗原对抗B IgM/IgG的吸附效率分别为96.88 %、98.44 %。结论：磁珠纯化抗原能特异性地与对应抗体结合，有效吸附血清中的血型抗体，有望作为合成A/B抗原的替代品。 |
| 英文摘要: |
| ABSTRACT Objective: To obtain A/B blood group antigen by immunomagnetic separation technique and explore whether the puri- fied antigens could be as an ABO blood group antibodies adsorbent to remove anti-A/B in serum. Methods: Magnetic beads coated with antibody were mixed with the pretreated saliva containing blood group materials to purity A/B antigen. The antigenicity and purity were confirmed by enzyme-linked immunosorbent assay and agglutination inhibition assay. The antibody adsorption effect of acquired anti- gens were evaluated by using unpurified antigens and purified antigens coated beads to adsorb antibodies in the serums which containing A/B antibodies, and using purified antigens coated beads to adsorb antibodies in the serums which from O-type pregnant woman. Results: The absorption value of wells adding purified antigen and the corresponding antibody were significantly high than the control wells (A antigen and anti-A 0.85±0.12 vs. 0.27±0.03, P<0.01; B antigen and anti-B 0.86±0.09 vs. 0.24±0.06, P<0.05), while the difference be- tween test well and control was not statistical significance when purified antigen reacted with other type antibodies. In the erythrocyte ag- glutination inhibition experiment, the antigen we purified could inhibit the reaction between the corresponding antibody and erythrocytes completely, but had no effect on the reaction of other type antibodies and erythrocytes. Then when we did serum antibody adsorption test, we found that purified antigen had higher adsorption efficiency than unpurified antigen (97.00 % vs. 88.00 %, P<0.001). In serum anti- body adsorption test of clinical samples, the adsorption efficiency of purified A antigens against anti-A IgG/IgM were 96.88 %, 98.44 % respectively, purified B antigen against anti-B IgM/IgG were 96.88 %, 98.44 % respectively. Conclusion: The purified antigens could specifically bind to anti-A or anti-B to adsorb blood group antibodies in serum with good effect, which were expected to be used as sub- stitutes for the synthetic A/B antigen. |
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