title: The preparation, characterization, and application of environment-friendly monoclonal antibodies for human blood cell.
abstract: OBJECTIVE
  Monoclonal anti-human blood group A (51A8) and B (63B6) antibody reagents were prepared using the serum-free technique. The aims of this research were to characterize the serum-free reagents and prove their reliabilities in routine use.
  
  
  METHODS
  Experiments including antigen-antibody agglutination testing, stability testing, SDS-PAGE, protein and IgM quantification, flow cytometry, and variable domain sequencing were performed to characterize the anti-A (51A8) and anti-B (63B6) reagents. Over 12 000 samples were tested using these reagents as routine blood grouping reagents.
  
  
  RESULTS
  Serum-free anti-A (51A8) and anti-B (63B6) reagents were stable in longitudinal and accelerated testing, and their high purity was shown in SDS-PAGE and IgM quantification. These reagents have high specificity to red blood cells in serologic agglutination testing and flow cytometric analysis. A1 and A2 subgroup antigens can be distinguished clearly by patterns of flow cytometric histograms. No discrepancy was found in clinical trials of 12 000 samples.
  
  
  CONCLUSIONS
  To reduce the risk of being affected by any animal additives, a serum-free culture system was applied to get mass-production of monoclonal anti-A/B antibodies. The high specificity and the high purity of the reagents were verified by the lab experiments.
  
  
  CONCLUSIONS
  Lab research and clinical trial showed that serum-free monoclonal anti-A (51A8) and anti-B (63B6) reagents meet the requirements of routine blood grouping reagents. Moreover, these reagents featured ultra-high purity that is missing in other commercial counterparts, and therefore are recommended as more environment-friendly reagents.
authors.affiliation: a Shanghai Biosci Biotechnology Co., Ltd , China.
authors.forename: Chenjie
authors.initials: C
authors.lastname: Zhou
authors.affiliation: a Shanghai Biosci Biotechnology Co., Ltd , China.
authors.forename: Xuechao
authors.initials: X
authors.lastname: Gao
authors.affiliation: a Shanghai Biosci Biotechnology Co., Ltd , China.
authors.forename: Shixiang
authors.initials: S
authors.lastname: He
authors.affiliation: a Shanghai Biosci Biotechnology Co., Ltd , China.
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Gao
authors.affiliation: a Shanghai Biosci Biotechnology Co., Ltd , China.
authors.forename: Jialin
authors.initials: J
authors.lastname: Zhuang
authors.affiliation: a Shanghai Biosci Biotechnology Co., Ltd , China.
authors.forename: Lirong
authors.initials: L
authors.lastname: Huang
authors.affiliation: a Shanghai Biosci Biotechnology Co., Ltd , China.
authors.forename: Hengchang
authors.initials: H
authors.lastname: Guo
mesh_terms: D000017:ABO Blood-Group System / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D004912:Erythrocytes / Q000276:immunology*; D006801:Humans; D051379:Mice
chemical_list: D000017:ABO Blood-Group System; D000911:Antibodies, Monoclonal
keywords: Anti-A; Anti-B; Monoclonal antibody; Serum-free