Skip to main content
Erschienen in: BMC Infectious Diseases 3/2014

Open Access 01.05.2014 | ePoster presentation

Production of cross neutralizing single chain fragment variables (scFv) from HIV-1 infected Indian children

verfasst von: Sanjeev Kumar, Rajesh Kumar, Muzamil Makhdoomi, Lubina Khan, SS Prakash, Ramachandran Thiruvengadam, Mohit Singla, Rakesh Lodha, SK Kabra, Subrata Sinha, Kalpana Luthra

Erschienen in: BMC Infectious Diseases | Sonderheft 3/2014

download
DOWNLOAD
print
DRUCKEN
insite
SUCHEN

Background

Monoclonal antibody based vaccines are effective and highly specific in disease targeting. Presently, most of the existing broadly neutralizing antibodies are generated against non subtype c viruses. HIV-1 subtype c accounts for more than 90% of infection in India. The disease progression in children is faster than adults.

Methods

Nine ART drug naïve HIV-1 subtype c infected children were recruited. PBMCs were isolated from all the subjects and pooled. RNA was isolated and cDNA was synthesized followed by amplification of VH and VL chain genes and scFv construction. A human recombinant scFv phage display library of 108 clones was constructed. Diversity of the phage library was checked by DNA sequencing and biopanned with RSC3 core antigen. 60 random clones were screened by phage ELISA. Expression of the scFvs was assessed by SDS-PAGE and Western blotting.

Results

The diversity of the phage library was more than 90%. Eight scFv monoclonals showed positive binding in phage ELISA and two best binders were further characterized. Both scFvs didn’t show any reactivity with unrelated antigens. DNA fingerprinting analysis showed both scFvs were distinct. A 32kDa band was obtained in SDS-PAGE and Western blot. Both scFvs showed cross neutralizing activity against a standard panel of pseudoviruses.

Conclusion

Here, we have for the first time generated a human recombinant scFv phage display library containing neutralizing clones from HIV-1 infected children. Further characterization of these scFvs generated against CD4 binding site for their epitope mapping would be helpful in the development of an effective HIV-1 vaccine.
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made.
The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.
The Creative Commons Public Domain Dedication waiver (https://​creativecommons.​org/​publicdomain/​zero/​1.​0/​) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
Metadaten
Titel
Production of cross neutralizing single chain fragment variables (scFv) from HIV-1 infected Indian children
verfasst von
Sanjeev Kumar
Rajesh Kumar
Muzamil Makhdoomi
Lubina Khan
SS Prakash
Ramachandran Thiruvengadam
Mohit Singla
Rakesh Lodha
SK Kabra
Subrata Sinha
Kalpana Luthra
Publikationsdatum
01.05.2014
Verlag
BioMed Central
Erschienen in
BMC Infectious Diseases / Ausgabe Sonderheft 3/2014
Elektronische ISSN: 1471-2334
DOI
https://doi.org/10.1186/1471-2334-14-S3-E25

Weitere Artikel der Sonderheft 3/2014

BMC Infectious Diseases 3/2014 Zur Ausgabe

Leitlinien kompakt für die Innere Medizin

Mit medbee Pocketcards sicher entscheiden.

Seit 2022 gehört die medbee GmbH zum Springer Medizin Verlag

Update Innere Medizin

Bestellen Sie unseren Fach-Newsletter und bleiben Sie gut informiert.