A cell sorter with modified bamboo charcoal for the efficient selection of specific antibody-producing hybridomas.
Monoclonal antibodies (mAbs) have been proven useful in research and clinical applications. However, the generation of mAbs by conventional hybridoma technology is time-, cost- and labor-consuming. Here we developed a simplified procedure for efficient generation and selection of antibody-producing hybridomas within 1 h, using a particular cell sorter design, a cytoflow reactor-based cell sorter (CBCS) which consists mainly of the "cytoflow reactor" that comprises two components, a reaction chamber and a glass tubing for air and medium exchange by gravity, and the "sorting material", human EGFR-conjugated bamboo charcoal, for specific B-cell enrichment. The high surface area and porous structure of bamboo charcoal greatly increased cell density and protein production. Moreover, from Raman, FT-IR spectroscopy and IFA analysis, the carboxylation and immobilization of bamboo charcoal can be introduced easily by nitric acid treatment and conjugated handily with human EGFR using EDC/NHS. Other evidences, such as IFA, showed that the specific hybridomas generated in this study could secrete specific anti-human EGFR antibodies. Our design allows the production of mAbs while avoiding time-consuming steps, such as large numbers of limiting dilutions and screening assays, and demonstrates that the CBCS could be a powerful tool for monoclonal antibody production.
Lin
Chien-Chen
CC
Institute of Biomedical Engineering, National Taiwan University, Taipei, Taiwan, ROC.
Ni
Mei-Hui
MH
Chang
Yu-Chung
YC
Yeh
Hsiu-Lun
HL
Lin
Feng-Huei
FH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000918:Antibody Specificity; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D019149:Bioreactors; D045744:Cell Line, Tumor; D002606:Charcoal / Q000737:chemistry* / Q000378:metabolism; D004867:Equipment Design; D066246:ErbB Receptors / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D055767:Immobilized Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008855:Microscopy, Electron, Scanning; D031787:Sasa / Q000737:chemistry; D013499:Surface Properties
D000911:Antibodies, Monoclonal; D055767:Immobilized Proteins; D002606:Charcoal; D066246:ErbB Receptors