![]() ![]() Two major strategies for generating fully human antibodies are: transgenic mice and antibody phage display. To date, 44 antibodies and antibody conjugates are EMA and/or FDA approved (status autumn 2014) ( ) and about 350 antibodies and antibody fusion proteins were under development in 2013. The main indications for therapeutic antibodies are cancer and auto-immune diseases. Therapeutic antibodies are currently one of the fastest developing class of biologicals in the pharmaceutical market. ![]() Since the inception of antibody technology twenty years ago, phage display is a powerful tool to generate antibodies for proteome research, diagnostics or for therapeutic purposes. Analysis of selected antibodies revealed that the complete amino acid diversity in the CDR-H3 was also found in selected scFvs showing the functionality of the naive CDR-H3 diversity. The highly diverse universal antibody gene libraries HAL9/10 were constructed using an optimized scFv phagemid vector design. Further, the data underline the importance of CDR length variations. The amino acid diversity and distribution in the CDR-H3 of the initial library was retrieved in the CDR-H3 of selected antibodies showing that all CDR-H3 amino acids occurring in the human antibody repertoire can be functionally used and is not biased by E. The HAL libraries and 834 antibodies selected against 121 targets were analyzed regarding the used germline V-genes, used V-gene combinations and CDR-H3/-元 length and composition. Deletion of a phenylalanine at the end of the CL linker sequence in our new phagemid design increased scFv production rate and frequency of selected kappa antibodies significantly. For several published antibody libraries, the selected number of kappa scFvs were lower compared to lambda scFvs, probably due to a lower kappa scFv or Fab expression rate. We show that changing the tag order to Myc/His improved the production of soluble antibodies, but did not affect antibody phage display. In detail, most common phagemids employed for antibody phage display are using a combined His/Myc tag for detection and purification. The naive antibody gene libraries HAL9 and HAL10, with a combined theoretical diversity of 1.5×10 10 independent clones, were constructed from 98 healthy donors using improved phage display vectors. Here we describe the design, construction and characterization of an optimized antibody phage display library. From naive antibody gene libraries - in theory - antibodies against any target can be selected. Antibody phage display is a proven key technology that allows the generation of human antibodies for diagnostics and therapy. ![]()
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