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In Vitro Evolution of Antibody Affinity using Libraries with Insertions and Deletions

repository. 2017; 
Kalliopi Skamaki
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GenParts™ DNA Fragments For pUC-N1-N2-MCS, pUC-g3p and pUC-N1-N2-HA vector construction, synthetic gene fragments of the pelB-N1-N2-MCS, pelB-MCS-gene III and HA peptide coding sequences flanked by appropriate restriction sites for cloning into the pUC19 vector were ordered from GenScript. Electrocompetent DH10B cells were used for cloning Get A Quote

摘要

In Nature, antibodies are capable of recognizing a huge variety of different molecular structures on the surface of antigens. The primary factor that defines the structural diversity of the antibody antigen combining site is the length variation of the complementarity determining region (CDR) loops. Following antigen stimulation, further diversification through the process called somatic hypermutation (SHM) leads to antibodies with improved affinity and specificity. Sequence diversification by SHM is mainly achieved by introduction of point substitutions and a small percentage of insertions/deletions (indels). Although the percentage of indels in affinity matured antibodies is low, probably due to the low rate ... More

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