Limits...
A high-affinity CDR-grafted antibody against influenza A H5N1 viruses recognizes a conserved epitope of H5 hemagglutinin.

Xiong F, Xia L, Wang J, Wu B, Wang D, Yuan L, Cheng Y, Zhu H, Che X, Zhang Q, Zhao G, Wang Y - PLoS ONE (2014)

Bottom Line: While vaccines and antiviral drugs are currently under development, neutralizing antibodies could offer an alternative strategy to prevent and treat H5N1 virus infection.Our results for the first time reported a humanized antibody against H5N1 viruses by CDR grafting method.With the expected lower immunogenicity, this humanized antibody was expected to be more efficacious than murine or human-mouse chimeric antibodies for future application in humans.

View Article: PubMed Central - PubMed

Affiliation: School of Life Science and Technology, Tongji University, Shanghai, China ; Shanghai-MOST Key Laboratory of Health and Disease Genomics, Chinese National Human Genome Center at Shanghai, Shanghai, China.

ABSTRACT
Highly pathogenic avian influenza (HPAI) H5N1 virus infection is still a potential threat to public health worldwide. While vaccines and antiviral drugs are currently under development, neutralizing antibodies could offer an alternative strategy to prevent and treat H5N1 virus infection. In the present study, we had developed a humanized antibody against H5N1 viruses from mouse-derived hybridoma in order to minimize its immunogenicity for potential clinical application. The humanized antibody hH5M9 was generated by transferring the mouse complementarity determining region (CDR) residues together with four key framework region (FR) residues onto the FR of the human antibody. This humanized antibody exhibited high affinity and specificity comparable to the parental mouse or chimeric counterpart with broad and strong neutralization activity against all H5N1 clades and subclades except for Egypt clades investigated. Furthermore, through epitope mapping we identified a linear epitope on the top region of hemagglutinin (HA) that was H5N1 specific and conserved. Our results for the first time reported a humanized antibody against H5N1 viruses by CDR grafting method. With the expected lower immunogenicity, this humanized antibody was expected to be more efficacious than murine or human-mouse chimeric antibodies for future application in humans.

Show MeSH

Related in: MedlinePlus

Characterization of cH5M9 and hH5M9 antibodies.(A) Antigen-binding capacity of mH5M9, cH5M9 and hH5M9 were measured by an indirect ELISA. (B) Affinity determination of mH5M9, cH5M9 and hH5M9 by surface plasmon resonance (SPR) assay. Various concentrations of mH5M9, cH5M9, hH5M9 and irrelevant anti-EGFR antibody C225 were injected on a GLC sensor chip immobilizing H5 HA (A/Vietnam/1194/04) for SPR kinetic binding analysis. Binding curves were recorded using ProteOn system in SPR biosensor instrument (BioRad Labs). (C) Calculated values of the association rate constant (Ka), dissociation rate constant (Kd), and equilibrium dissociation constant (KD) of mH5M9, cH5M9 and hH5M9.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3928294&req=5

pone-0088777-g002: Characterization of cH5M9 and hH5M9 antibodies.(A) Antigen-binding capacity of mH5M9, cH5M9 and hH5M9 were measured by an indirect ELISA. (B) Affinity determination of mH5M9, cH5M9 and hH5M9 by surface plasmon resonance (SPR) assay. Various concentrations of mH5M9, cH5M9, hH5M9 and irrelevant anti-EGFR antibody C225 were injected on a GLC sensor chip immobilizing H5 HA (A/Vietnam/1194/04) for SPR kinetic binding analysis. Binding curves were recorded using ProteOn system in SPR biosensor instrument (BioRad Labs). (C) Calculated values of the association rate constant (Ka), dissociation rate constant (Kd), and equilibrium dissociation constant (KD) of mH5M9, cH5M9 and hH5M9.

Mentions: An indirect ELISA was first used to analyze the antigen-binding activity of cH5M9 and hH5M9 against H5 HA (A/Vietnam/1194/04). As shown in Figure 2A, cH5M9 had the similar antigen-binding activity to mH5M9, while the antigen-binding activity of hH5M9 was lower than that of mH5M9, suggesting that the humanization of recombinant antibody by CDR grafting might result in the structural alteration of the antigen-binding site more or less and the subsequent decrease of antigen-binding capacity [32].


A high-affinity CDR-grafted antibody against influenza A H5N1 viruses recognizes a conserved epitope of H5 hemagglutinin.

Xiong F, Xia L, Wang J, Wu B, Wang D, Yuan L, Cheng Y, Zhu H, Che X, Zhang Q, Zhao G, Wang Y - PLoS ONE (2014)

Characterization of cH5M9 and hH5M9 antibodies.(A) Antigen-binding capacity of mH5M9, cH5M9 and hH5M9 were measured by an indirect ELISA. (B) Affinity determination of mH5M9, cH5M9 and hH5M9 by surface plasmon resonance (SPR) assay. Various concentrations of mH5M9, cH5M9, hH5M9 and irrelevant anti-EGFR antibody C225 were injected on a GLC sensor chip immobilizing H5 HA (A/Vietnam/1194/04) for SPR kinetic binding analysis. Binding curves were recorded using ProteOn system in SPR biosensor instrument (BioRad Labs). (C) Calculated values of the association rate constant (Ka), dissociation rate constant (Kd), and equilibrium dissociation constant (KD) of mH5M9, cH5M9 and hH5M9.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3928294&req=5

pone-0088777-g002: Characterization of cH5M9 and hH5M9 antibodies.(A) Antigen-binding capacity of mH5M9, cH5M9 and hH5M9 were measured by an indirect ELISA. (B) Affinity determination of mH5M9, cH5M9 and hH5M9 by surface plasmon resonance (SPR) assay. Various concentrations of mH5M9, cH5M9, hH5M9 and irrelevant anti-EGFR antibody C225 were injected on a GLC sensor chip immobilizing H5 HA (A/Vietnam/1194/04) for SPR kinetic binding analysis. Binding curves were recorded using ProteOn system in SPR biosensor instrument (BioRad Labs). (C) Calculated values of the association rate constant (Ka), dissociation rate constant (Kd), and equilibrium dissociation constant (KD) of mH5M9, cH5M9 and hH5M9.
Mentions: An indirect ELISA was first used to analyze the antigen-binding activity of cH5M9 and hH5M9 against H5 HA (A/Vietnam/1194/04). As shown in Figure 2A, cH5M9 had the similar antigen-binding activity to mH5M9, while the antigen-binding activity of hH5M9 was lower than that of mH5M9, suggesting that the humanization of recombinant antibody by CDR grafting might result in the structural alteration of the antigen-binding site more or less and the subsequent decrease of antigen-binding capacity [32].

Bottom Line: While vaccines and antiviral drugs are currently under development, neutralizing antibodies could offer an alternative strategy to prevent and treat H5N1 virus infection.Our results for the first time reported a humanized antibody against H5N1 viruses by CDR grafting method.With the expected lower immunogenicity, this humanized antibody was expected to be more efficacious than murine or human-mouse chimeric antibodies for future application in humans.

View Article: PubMed Central - PubMed

Affiliation: School of Life Science and Technology, Tongji University, Shanghai, China ; Shanghai-MOST Key Laboratory of Health and Disease Genomics, Chinese National Human Genome Center at Shanghai, Shanghai, China.

ABSTRACT
Highly pathogenic avian influenza (HPAI) H5N1 virus infection is still a potential threat to public health worldwide. While vaccines and antiviral drugs are currently under development, neutralizing antibodies could offer an alternative strategy to prevent and treat H5N1 virus infection. In the present study, we had developed a humanized antibody against H5N1 viruses from mouse-derived hybridoma in order to minimize its immunogenicity for potential clinical application. The humanized antibody hH5M9 was generated by transferring the mouse complementarity determining region (CDR) residues together with four key framework region (FR) residues onto the FR of the human antibody. This humanized antibody exhibited high affinity and specificity comparable to the parental mouse or chimeric counterpart with broad and strong neutralization activity against all H5N1 clades and subclades except for Egypt clades investigated. Furthermore, through epitope mapping we identified a linear epitope on the top region of hemagglutinin (HA) that was H5N1 specific and conserved. Our results for the first time reported a humanized antibody against H5N1 viruses by CDR grafting method. With the expected lower immunogenicity, this humanized antibody was expected to be more efficacious than murine or human-mouse chimeric antibodies for future application in humans.

Show MeSH
Related in: MedlinePlus