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Bifunctional recombinant fusion proteins for rapid detection of antibodies to both HIV-1 and HIV-2 in whole blood

BACKGROUND: Availability of accurate diagnostic tests has been helpful in curtailing the spread of HIV infection. Among these, simple, point of care, inexpensive tests which require only a drop of blood from finger-prick and give reliable results within minutes are a must for expansion of testing se...

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Autores principales: Gupta, Amita, Chaudhary, Vijay K
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2006
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1599719/
https://www.ncbi.nlm.nih.gov/pubmed/16995928
http://dx.doi.org/10.1186/1472-6750-6-39
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author Gupta, Amita
Chaudhary, Vijay K
author_facet Gupta, Amita
Chaudhary, Vijay K
author_sort Gupta, Amita
collection PubMed
description BACKGROUND: Availability of accurate diagnostic tests has been helpful in curtailing the spread of HIV infection. Among these, simple, point of care, inexpensive tests which require only a drop of blood from finger-prick and give reliable results within minutes are a must for expansion of testing services and for reaching mobile and marginalised populations. Such tests will not only be a boon for the infrastructure-starved developing and underdeveloped countries but will also be extremely useful in developed countries where post-testing compliance is a major problem. Our laboratory has been involved in developing reagents for heamagglutination-based rapid detection of antibodies to HIV in whole blood using recombinant molecules specific for either HIV-1 or HIV-2. Since it is not required of a screening test to differentially detect HIV and HIV-2, it would useful to create a single molecule capable of simultaneous detection of both HIV-1 and HIV-2 in a drop of blood. RESULTS: The present paper describes designing, high-level expression and large-scale purification of new molecules comprising recombinant anti-RBC Fab fused to immunodominant regions of envelope sequences from both gp41 of HIV-1 and gp36 of HIV-2. These immunodominant regions of HIV envelope contain cysteine residues, which make disulfide bond and can interfere with the assembly of light chain and heavy chain fragment to make Fab molecule in vitro. To circumvent this problem, a series of fusion proteins having different combinations of native and mutant envelope sequences were constructed, purified and evaluated for their efficacy in detecting antibodies to HIV-1 and HIV-2. A chimeric molecule comprising native envelope sequence of gp41 of HIV-1 and modified envelope sequence of gp36 of HIV-2 gave good production yield and also detected both HIV-1 and HIV-2 samples with high sensitivity and specificity. CONCLUSION: The new bifunctional antibody fusion protein identified in this study detects both HIV-1 and HIV-2 infected samples efficiently and can be used in place of molecules that detect only HIV-1 or HIV-2. This will make reagent production more economical as only one molecule has to be produced in place of two molecules. Also, it will simplify the testing procedure allowing detection of both HIV-1 and HIV-2 infections in a single drop of blood.
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spelling pubmed-15997192006-10-12 Bifunctional recombinant fusion proteins for rapid detection of antibodies to both HIV-1 and HIV-2 in whole blood Gupta, Amita Chaudhary, Vijay K BMC Biotechnol Research Article BACKGROUND: Availability of accurate diagnostic tests has been helpful in curtailing the spread of HIV infection. Among these, simple, point of care, inexpensive tests which require only a drop of blood from finger-prick and give reliable results within minutes are a must for expansion of testing services and for reaching mobile and marginalised populations. Such tests will not only be a boon for the infrastructure-starved developing and underdeveloped countries but will also be extremely useful in developed countries where post-testing compliance is a major problem. Our laboratory has been involved in developing reagents for heamagglutination-based rapid detection of antibodies to HIV in whole blood using recombinant molecules specific for either HIV-1 or HIV-2. Since it is not required of a screening test to differentially detect HIV and HIV-2, it would useful to create a single molecule capable of simultaneous detection of both HIV-1 and HIV-2 in a drop of blood. RESULTS: The present paper describes designing, high-level expression and large-scale purification of new molecules comprising recombinant anti-RBC Fab fused to immunodominant regions of envelope sequences from both gp41 of HIV-1 and gp36 of HIV-2. These immunodominant regions of HIV envelope contain cysteine residues, which make disulfide bond and can interfere with the assembly of light chain and heavy chain fragment to make Fab molecule in vitro. To circumvent this problem, a series of fusion proteins having different combinations of native and mutant envelope sequences were constructed, purified and evaluated for their efficacy in detecting antibodies to HIV-1 and HIV-2. A chimeric molecule comprising native envelope sequence of gp41 of HIV-1 and modified envelope sequence of gp36 of HIV-2 gave good production yield and also detected both HIV-1 and HIV-2 samples with high sensitivity and specificity. CONCLUSION: The new bifunctional antibody fusion protein identified in this study detects both HIV-1 and HIV-2 infected samples efficiently and can be used in place of molecules that detect only HIV-1 or HIV-2. This will make reagent production more economical as only one molecule has to be produced in place of two molecules. Also, it will simplify the testing procedure allowing detection of both HIV-1 and HIV-2 infections in a single drop of blood. BioMed Central 2006-09-22 /pmc/articles/PMC1599719/ /pubmed/16995928 http://dx.doi.org/10.1186/1472-6750-6-39 Text en Copyright © 2006 Gupta and Chaudhary; 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 Research Article
Gupta, Amita
Chaudhary, Vijay K
Bifunctional recombinant fusion proteins for rapid detection of antibodies to both HIV-1 and HIV-2 in whole blood
title Bifunctional recombinant fusion proteins for rapid detection of antibodies to both HIV-1 and HIV-2 in whole blood
title_full Bifunctional recombinant fusion proteins for rapid detection of antibodies to both HIV-1 and HIV-2 in whole blood
title_fullStr Bifunctional recombinant fusion proteins for rapid detection of antibodies to both HIV-1 and HIV-2 in whole blood
title_full_unstemmed Bifunctional recombinant fusion proteins for rapid detection of antibodies to both HIV-1 and HIV-2 in whole blood
title_short Bifunctional recombinant fusion proteins for rapid detection of antibodies to both HIV-1 and HIV-2 in whole blood
title_sort bifunctional recombinant fusion proteins for rapid detection of antibodies to both hiv-1 and hiv-2 in whole blood
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1599719/
https://www.ncbi.nlm.nih.gov/pubmed/16995928
http://dx.doi.org/10.1186/1472-6750-6-39
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