Emanuel P, O'Brien T, Burans J, DasGupta B R, Valdes J J, Eldefrawi M
Department of Pharmacology and Experimental Therapeutics, School of Medicine, University of Maryland, Baltimore 21201, USA.
J Immunol Methods. 1996 Jun 21;193(2):189-97. doi: 10.1016/0022-1759(96)00053-1.
The production of antibodies towards antigens with low immunogenicity is enhanced by the intrinsic efficiency of screening combinatorial libraries of immunoglobulins. The need to isolate clones with rare binding specificities has dictated a highly efficient method of screening and isolating antibody clones. The production of recombinant immunoglobulin libraries in bacteria allows for a more controlled selection of antibody specificity and can be used in circumstances where hybridoma fusions are unable to isolate rare clones with the desired epitope specificity. Botulinum neurotoxin (NT) with associated non neurotoxin proteins (non-NT) as a complex was used to immunize mice to obtain mRNA for the production of a recombinant antibody library with a repertoire of specificities. Initial screens of the combinatorial library revealed clones which recognized the non-neurotoxin proteins of the toxin complex similar to monoclonal antibodies produced by conventional hybridoma fusions. The combinatorial library was re-screened in order to isolate antibodies that specifically recognized the neurotoxin component of the toxin complex. The ability to alter the biopanning selection process affords the researcher a measure of control in the selection process not available with traditional hybridoma fusions.
通过筛选免疫球蛋白组合文库的内在效率，可增强针对低免疫原性抗原的抗体产生。分离具有罕见结合特异性克隆的需求促使了一种高效筛选和分离抗体克隆的方法的产生。在细菌中生产重组免疫球蛋白文库能够更可控地选择抗体特异性，并且可用于杂交瘤融合无法分离出具有所需表位特异性的罕见克隆的情况。将肉毒杆菌神经毒素（NT）与相关非神经毒素蛋白（非NT）作为复合物用于免疫小鼠，以获取用于生产具有多种特异性的重组抗体文库的mRNA。对组合文库的初步筛选揭示了一些克隆，这些克隆识别毒素复合物的非神经毒素蛋白，类似于传统杂交瘤融合产生的单克隆抗体。对组合文库进行重新筛选，以分离出特异性识别毒素复合物神经毒素成分的抗体。改变淘选选择过程的能力使研究人员在选择过程中具有一定程度的控制权，这是传统杂交瘤融合所不具备的。
