Cargando…

Evolutional selection of a combinatorial phage library displaying randomly-rearranged various single domains of immunoglobulin (Ig)-binding proteins (IBPs) with four kinds of Ig molecules

BACKGROUND: Protein A, protein G and protein L are three well-defined immunoglobulin (Ig)-binding proteins (IBPs), which show affinity for specific sites on Ig of mammalian hosts. Although the precise functions of these molecules are not fully understood, it is thought that they play an important ro...

Descripción completa

Detalles Bibliográficos
Autores principales: Yang, Hua, Cao, Jie, Li, Lian-Qing, Zhou, Xia, Chen, Qiu-Li, Liao, Wen-Ting, Wen, Zong-Mei, Jiang, Shao-Hua, Xu, Rong, Jia, Jian-An, Pan, Xin, Qi, Zhong-Tian, Pan, Wei
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2532689/
https://www.ncbi.nlm.nih.gov/pubmed/18700046
http://dx.doi.org/10.1186/1471-2180-8-137
_version_ 1782158988004032512
author Yang, Hua
Cao, Jie
Li, Lian-Qing
Zhou, Xia
Chen, Qiu-Li
Liao, Wen-Ting
Wen, Zong-Mei
Jiang, Shao-Hua
Xu, Rong
Jia, Jian-An
Pan, Xin
Qi, Zhong-Tian
Pan, Wei
author_facet Yang, Hua
Cao, Jie
Li, Lian-Qing
Zhou, Xia
Chen, Qiu-Li
Liao, Wen-Ting
Wen, Zong-Mei
Jiang, Shao-Hua
Xu, Rong
Jia, Jian-An
Pan, Xin
Qi, Zhong-Tian
Pan, Wei
author_sort Yang, Hua
collection PubMed
description BACKGROUND: Protein A, protein G and protein L are three well-defined immunoglobulin (Ig)-binding proteins (IBPs), which show affinity for specific sites on Ig of mammalian hosts. Although the precise functions of these molecules are not fully understood, it is thought that they play an important role in pathogenicity of bacteria. The single domains of protein A, protein G and protein L were all demonstrated to have function to bind to Ig. Whether combinations of Ig-binding domains of various IBPs could exhibit useful novel binding is interesting. RESULTS: We used a combinatorial phage library which displayed randomly-rearranged various-peptide-linked molecules of D and A domains of protein A, designated PA(D) and PA(A) respectively, B2 domain of protein G (PG) and B3 domain of protein L (PL) for affinity selection with human IgG (hIgG), human IgM (hIgM), human IgA (hIgA) and recombinant hIgG1-Fc as bait respectively. Two kinds of novel combinatorial molecules with characteristic structure of PA(A)-PG and PA(A)-PL were obtained in hIgG (hIgG1-Fc) and hIgM (hIgA) post-selection populations respectively. In addition, the linking peptides among all PA(A)-PG and PA(A)-PL structures was strongly selected, and showed interestingly divergent and convergent distribution. The phage binding assays and competitive inhibition experiments demonstrated that PA(A)-PG and PA(A)-PL combinations possess comparable binding advantages with hIgG/hIgG1-Fc and hIgM/hIgA respectively. CONCLUSION: In this work, a combinatorial phage library displaying Ig-binding domains of protein A, protein G, or protein L joined by various random linking peptides was used to conducted evolutional selection in vitro with four kinds of Ig molecules. Two kinds of novel combinations of Ig-binding domains, PA(A)-PG and PA(A)-PL, were obtained, and demonstrate the novel Ig binding properties.
format Text
id pubmed-2532689
institution National Center for Biotechnology Information
language English
publishDate 2008
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-25326892008-09-09 Evolutional selection of a combinatorial phage library displaying randomly-rearranged various single domains of immunoglobulin (Ig)-binding proteins (IBPs) with four kinds of Ig molecules Yang, Hua Cao, Jie Li, Lian-Qing Zhou, Xia Chen, Qiu-Li Liao, Wen-Ting Wen, Zong-Mei Jiang, Shao-Hua Xu, Rong Jia, Jian-An Pan, Xin Qi, Zhong-Tian Pan, Wei BMC Microbiol Methodology Article BACKGROUND: Protein A, protein G and protein L are three well-defined immunoglobulin (Ig)-binding proteins (IBPs), which show affinity for specific sites on Ig of mammalian hosts. Although the precise functions of these molecules are not fully understood, it is thought that they play an important role in pathogenicity of bacteria. The single domains of protein A, protein G and protein L were all demonstrated to have function to bind to Ig. Whether combinations of Ig-binding domains of various IBPs could exhibit useful novel binding is interesting. RESULTS: We used a combinatorial phage library which displayed randomly-rearranged various-peptide-linked molecules of D and A domains of protein A, designated PA(D) and PA(A) respectively, B2 domain of protein G (PG) and B3 domain of protein L (PL) for affinity selection with human IgG (hIgG), human IgM (hIgM), human IgA (hIgA) and recombinant hIgG1-Fc as bait respectively. Two kinds of novel combinatorial molecules with characteristic structure of PA(A)-PG and PA(A)-PL were obtained in hIgG (hIgG1-Fc) and hIgM (hIgA) post-selection populations respectively. In addition, the linking peptides among all PA(A)-PG and PA(A)-PL structures was strongly selected, and showed interestingly divergent and convergent distribution. The phage binding assays and competitive inhibition experiments demonstrated that PA(A)-PG and PA(A)-PL combinations possess comparable binding advantages with hIgG/hIgG1-Fc and hIgM/hIgA respectively. CONCLUSION: In this work, a combinatorial phage library displaying Ig-binding domains of protein A, protein G, or protein L joined by various random linking peptides was used to conducted evolutional selection in vitro with four kinds of Ig molecules. Two kinds of novel combinations of Ig-binding domains, PA(A)-PG and PA(A)-PL, were obtained, and demonstrate the novel Ig binding properties. BioMed Central 2008-08-13 /pmc/articles/PMC2532689/ /pubmed/18700046 http://dx.doi.org/10.1186/1471-2180-8-137 Text en Copyright © 2008 Yang et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methodology Article
Yang, Hua
Cao, Jie
Li, Lian-Qing
Zhou, Xia
Chen, Qiu-Li
Liao, Wen-Ting
Wen, Zong-Mei
Jiang, Shao-Hua
Xu, Rong
Jia, Jian-An
Pan, Xin
Qi, Zhong-Tian
Pan, Wei
Evolutional selection of a combinatorial phage library displaying randomly-rearranged various single domains of immunoglobulin (Ig)-binding proteins (IBPs) with four kinds of Ig molecules
title Evolutional selection of a combinatorial phage library displaying randomly-rearranged various single domains of immunoglobulin (Ig)-binding proteins (IBPs) with four kinds of Ig molecules
title_full Evolutional selection of a combinatorial phage library displaying randomly-rearranged various single domains of immunoglobulin (Ig)-binding proteins (IBPs) with four kinds of Ig molecules
title_fullStr Evolutional selection of a combinatorial phage library displaying randomly-rearranged various single domains of immunoglobulin (Ig)-binding proteins (IBPs) with four kinds of Ig molecules
title_full_unstemmed Evolutional selection of a combinatorial phage library displaying randomly-rearranged various single domains of immunoglobulin (Ig)-binding proteins (IBPs) with four kinds of Ig molecules
title_short Evolutional selection of a combinatorial phage library displaying randomly-rearranged various single domains of immunoglobulin (Ig)-binding proteins (IBPs) with four kinds of Ig molecules
title_sort evolutional selection of a combinatorial phage library displaying randomly-rearranged various single domains of immunoglobulin (ig)-binding proteins (ibps) with four kinds of ig molecules
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2532689/
https://www.ncbi.nlm.nih.gov/pubmed/18700046
http://dx.doi.org/10.1186/1471-2180-8-137
work_keys_str_mv AT yanghua evolutionalselectionofacombinatorialphagelibrarydisplayingrandomlyrearrangedvarioussingledomainsofimmunoglobulinigbindingproteinsibpswithfourkindsofigmolecules
AT caojie evolutionalselectionofacombinatorialphagelibrarydisplayingrandomlyrearrangedvarioussingledomainsofimmunoglobulinigbindingproteinsibpswithfourkindsofigmolecules
AT lilianqing evolutionalselectionofacombinatorialphagelibrarydisplayingrandomlyrearrangedvarioussingledomainsofimmunoglobulinigbindingproteinsibpswithfourkindsofigmolecules
AT zhouxia evolutionalselectionofacombinatorialphagelibrarydisplayingrandomlyrearrangedvarioussingledomainsofimmunoglobulinigbindingproteinsibpswithfourkindsofigmolecules
AT chenqiuli evolutionalselectionofacombinatorialphagelibrarydisplayingrandomlyrearrangedvarioussingledomainsofimmunoglobulinigbindingproteinsibpswithfourkindsofigmolecules
AT liaowenting evolutionalselectionofacombinatorialphagelibrarydisplayingrandomlyrearrangedvarioussingledomainsofimmunoglobulinigbindingproteinsibpswithfourkindsofigmolecules
AT wenzongmei evolutionalselectionofacombinatorialphagelibrarydisplayingrandomlyrearrangedvarioussingledomainsofimmunoglobulinigbindingproteinsibpswithfourkindsofigmolecules
AT jiangshaohua evolutionalselectionofacombinatorialphagelibrarydisplayingrandomlyrearrangedvarioussingledomainsofimmunoglobulinigbindingproteinsibpswithfourkindsofigmolecules
AT xurong evolutionalselectionofacombinatorialphagelibrarydisplayingrandomlyrearrangedvarioussingledomainsofimmunoglobulinigbindingproteinsibpswithfourkindsofigmolecules
AT jiajianan evolutionalselectionofacombinatorialphagelibrarydisplayingrandomlyrearrangedvarioussingledomainsofimmunoglobulinigbindingproteinsibpswithfourkindsofigmolecules
AT panxin evolutionalselectionofacombinatorialphagelibrarydisplayingrandomlyrearrangedvarioussingledomainsofimmunoglobulinigbindingproteinsibpswithfourkindsofigmolecules
AT qizhongtian evolutionalselectionofacombinatorialphagelibrarydisplayingrandomlyrearrangedvarioussingledomainsofimmunoglobulinigbindingproteinsibpswithfourkindsofigmolecules
AT panwei evolutionalselectionofacombinatorialphagelibrarydisplayingrandomlyrearrangedvarioussingledomainsofimmunoglobulinigbindingproteinsibpswithfourkindsofigmolecules