Cross-neutralizing anti-HIV-1 human single chain variable fragments(scFvs) against CD4 binding site and N332 glycan identified from a recombinant phage library.
More than 50% of HIV-1 infection globally is caused by subtype_C viruses. Majority of the broadly neutralizing antibodies (bnAbs) targeting HIV-1 have been isolated from non-subtype_C infected donors. Mapping the epitope specificities of bnAbs provides useful information for vaccine design. Recombinant antibody technology enables generation of a large repertoire of monoclonals with diverse specificities. We constructed a phage recombinant single chain variable fragment (scFv) library with a diversity of 7.8 × 108clones, using a novel strategy of pooling peripheral blood mononuclear cells (PBMCs) of six select HIV-1 chronically infected Indian donors whose plasma antibodies exhibited potent cross neutralization efficiency. The library was panned and screened by phage ELISA using trimeric recombinant proteins to identify viral envelope specific clones. Three scFv monoclonals D11, C11 and 1F6 selected from the library cross neutralized subtypes A, B and C viruses at concentrations ranging from 0.09 μg/mL to 100 μg/mL. The D11 and 1F6 scFvs competed with mAbs b12 and VRC01 demonstrating CD4bs specificity, while C11 demonstrated N332 specificity. This is the first study to identify cross neutralizing scFv monoclonals with CD4bs and N332 glycan specificities from India. Cross neutralizing anti-HIV-1 human scFv monoclonals can be potential candidates for passive immunotherapy and for guiding immunogen design.
Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, India.
Lubina
L
Khan
Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, India.
Rajesh
R
Kumar
Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, India.
Ramachandran
R
Thiruvengadam
Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, India.
Hilal Ahmad
HA
Parray
Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, India.
Muzamil Ashraf
MA
Makhdoomi
Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, India.
Sanjeev
S
Kumar
Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, India.
Heena
H
Aggarwal
Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, India.
Madhav
M
Mohata
Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, India.
Abdul Wahid
AW
Hussain
Molecular Biophysics Unit, Indian Institute of Science, Bangalore, India.
Raksha
R
Das
Molecular Biophysics Unit, Indian Institute of Science, Bangalore, India.
Raghavan
R
Varadarajan
HIV Vaccine Translational Research Laboratory, Translational Health Science and Technology Institute, NCR Biotech Science Cluster, Faridabad, Haryana, India.|International AIDS Vaccine initiative, USA.
Jayanta
J
Bhattacharya
Department of Microbiology, All India Institute of Medical Sciences, New Delhi, India.
Madhu
M
Vajpayee
Y.R. Gaitonde Centre for AIDS Research and Education (YRG CARE), Chennai, India.
K G
KG
Murugavel
Y.R. Gaitonde Centre for AIDS Research and Education (YRG CARE), Chennai, India.
Suniti
S
Solomon
Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, India.|National Brain Research Centre, Manesar, Gurgaon, Haryana, India.
Subrata
S
Sinha
Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, India.
Kalpana
K
Luthra
D057134:Antibodies, Neutralizing / Q000276:immunology; D000915:Antibody Affinity; D001665:Binding Sites / Q000276:immunology*; D015704:CD4 Antigens / Q000378:metabolism*; D018604:Epitope Mapping; D015483:HIV Antibodies / Q000235:genetics / Q000276:immunology*; D015699:HIV Envelope Protein gp120 / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D015658:HIV Infections / Q000276:immunology* / Q000821:virology; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D009500:Neutralization Tests; D019151:Peptide Library; D011485:Protein Binding / Q000276:immunology; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology*
D057134:Antibodies, Neutralizing; D015704:CD4 Antigens; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D019151:Peptide Library; D057127:Single-Chain Antibodies