Improved cell line development by a high throughput affinity capture surface display technique to select for high secretors.
A novel process is described which permits rapid and objective selection of rare cells from a heterogeneous population based on quantity of secreted target protein. The process involves construction of an immobilised affinity surface display matrix that specifically binds secreted target product which is then detected using a fluorescent labelled ligand. Cells with the highest fluorescence can then be sorted using conventional flow cytometric technology. Overall, the whole process can be completed in less than 4 h during which time in the region of five million cells can be analysed. Cells are rapidly selected for in a quantitative manner compared to traditional methods which can take several months and have a reduced probability of finding low abundance high secretors due to practical limitations imposed on the number of cells which can be screened.
Holmes
P
P
Animal Cell Technology Group, School of Chemical Engineering, The University of Birmingham, Edgbaston, Birmingham, UK.
Al-Rubeai
M
M
D000345:Affinity Labels; D000818:Animals; D001360:Avidin / Q000378:metabolism; D001710:Biotin / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D002469:Cell Separation; D005069:Evaluation Studies as Topic; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D007158:Immunologic Techniques; D007700:Kinetics; D051379:Mice; D009101:Multiple Myeloma / Q000276:immunology / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000345:Affinity Labels; D005456:Fluorescent Dyes; D011506:Proteins; D011956:Receptors, Cell Surface; D001360:Avidin; D001710:Biotin