Selection and affinity maturation of human antibodies against rabies virus from a scFv gene library using ribosome display.
We selected useful antibody fragments against rabies virus from a human single chain variable fragment (scFv) gene library using ribosome display technique. The recombinant rabies virus glycoprotein (RVGp) was used as an antigen to isolate specific scFvs. After five rounds of selection, the analysis demonstrated that scFv-ribosome-mRNA complexes were specifically selected against RVGp. Sequence analysis showed that mutations were introduced at random by PCR among the rounds of selection and variants with high affinity were isolated. The obtained scFvs with high affinity could recognize RVGp specifically and showed binding activity to rabies virus. These scFvs were potential for inclusion in a combination of several human monoclonal antibodies (MAbs) aimed for application in rabies post-exposure prophylaxis. Ribosome display technology is a robust tool for rapid isolation of human antibody fragments, and has exceptional strength in affinity maturation and molecular evolution in vitro.
Zhao
Xiao-Ling
XL
Institute of Health and Environmental Medicine, Academy of Military Medical Sciences, Heping District, Tianjin 300050, China. xlzhao985@sohu.com
Chen
Wei-Qiang
WQ
Yang
Zhi-Hua
ZH
Li
Jing-Mei
JM
Zhang
Su-Juan
SJ
Tian
Lu-Fang
LF
D000914:Antibodies, Viral / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D000915:Antibody Affinity; D001709:Biotechnology / Q000379:methods; D015723:Gene Library; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011820:Rabies virus / Q000276:immunology*; D012270:Ribosomes / Q000378:metabolism*; D012641:Selection, Genetic; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology
D000914:Antibodies, Viral; D006023:Glycoproteins; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D012333:RNA, Messenger; D014764:Viral Proteins