In vitro antibody affinity maturation targeting germline hotspots.

In vitro antibody affinity maturation targeting germline hotspots.
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DOI:
10.1007/978-1-59745-554-1_15
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发表时间:
2009
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
--
通讯作者:
Pastan, Ira
Pastan, Ira
中科院分区:
其他
文献类型:
--
作者:
Ho, Mitchell;Pastan, Ira

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Affinity-matured antibodies can exhibit increased biological efficacy. Regardless of whether an antibody is isolated from a hybridoma or a human Fv phage library, the antibody affinity for its target may need improvement for therapeutic applications. An increased affinity may allow for a reduced dosage of a therapeutic antibody; toxic side effects may also be reduced. In the immune system, affinity maturation is a process involving somatic hypermutations in B cells. Therefore, germline hotspot residues are most likely to have a major impact on antibody affinity. Here, we describe procedures for germline hotspot mutagenesis with an emphasis on strategies for randomizing hotspots with PCR and phage display, using as an example the anti-CD22 monoclonal antibody.
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