Isolation, Characterization, And Production Of Recombinant Monoclonal Antibodies Against Rnie For Development Of A Strongyloides Antigen Detection Assay

Strongyloides stercoralis is a soil-transmitted helminth that causes strongyloidiasis. It is estimated to infect more than 600 million people worldwide. Asymptomatic chronic infections in immunocompromised people can lead to fatal hyperinfection. Serodiagnosis by detecting specific IgG antibodies ca...

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Main Author:	Balachandra, Dinesh
Format:	Thesis
Language:	English
Published:	2022
Subjects:	R5-920 Medicine (General)
Online Access:	http://eprints.usm.my/59388/1/DINESH%20AL%20BALACHANDRA%20-%20TESIS24.pdf
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id	my-usm-ep.59388
record_format	uketd_dc
spelling	my-usm-ep.593882023-09-14T08:41:39Z Isolation, Characterization, And Production Of Recombinant Monoclonal Antibodies Against Rnie For Development Of A Strongyloides Antigen Detection Assay 2022-05 Balachandra, Dinesh R5-920 Medicine (General) Strongyloides stercoralis is a soil-transmitted helminth that causes strongyloidiasis. It is estimated to infect more than 600 million people worldwide. Asymptomatic chronic infections in immunocompromised people can lead to fatal hyperinfection. Serodiagnosis by detecting specific IgG antibodies can be challenging due to potential cross-reactivity with infections by other parasites. An antigen detection assay, a direct detection method, can help the diagnosis and is useful for post-treatment follow-up. This study used phage display technology to produce recombinant monoclonal antibodies (rMAb) against NIE recombinant protein (rNIE) and develop a Strongyloides antigen detection test. rNIE is an established protein for the diagnosis of strongyloidiasis. rNIE was expressed, purified, and then used to select rMAb candidates via biopanning of an immune helminth phage display library. It isolated of 104 ELISA-positive clones and sequence analysis showed that 30 clones had full-length light and heavy chains. Four unique gene families were identified, i.e., IgHV3-LV6 (86.66%), IgHV1-LV3 (3.33%), IgHV5-KV3 (3.33%), and IgHV3-LV3 (6.66%). Randomly, one representative clone from each gene family was selected for further studies, i.e., (a) rMAb5 representing IgHV1-LV3, (b) rMAb6 representing IgHV3-LV6, (c) rMAb14 representing IgHV5-KV3, and (d) rMAb23 representing IgHV3-LV3. The rMAb gene sequences from the phage display vector were subcloned into the pET51b+ expression vector and transformed into Escherichia coli Shuffle T7 Express host cell. 2022-05 Thesis http://eprints.usm.my/59388/ http://eprints.usm.my/59388/1/DINESH%20AL%20BALACHANDRA%20-%20TESIS24.pdf application/pdf en public phd doctoral Universiti Sains Malaysia Institut Penyelidikan Perubatan Molekul
institution	Universiti Sains Malaysia
collection	USM Institutional Repository
language	English
topic	R5-920 Medicine (General)
spellingShingle	R5-920 Medicine (General) Balachandra, Dinesh Isolation, Characterization, And Production Of Recombinant Monoclonal Antibodies Against Rnie For Development Of A Strongyloides Antigen Detection Assay
description	Strongyloides stercoralis is a soil-transmitted helminth that causes strongyloidiasis. It is estimated to infect more than 600 million people worldwide. Asymptomatic chronic infections in immunocompromised people can lead to fatal hyperinfection. Serodiagnosis by detecting specific IgG antibodies can be challenging due to potential cross-reactivity with infections by other parasites. An antigen detection assay, a direct detection method, can help the diagnosis and is useful for post-treatment follow-up. This study used phage display technology to produce recombinant monoclonal antibodies (rMAb) against NIE recombinant protein (rNIE) and develop a Strongyloides antigen detection test. rNIE is an established protein for the diagnosis of strongyloidiasis. rNIE was expressed, purified, and then used to select rMAb candidates via biopanning of an immune helminth phage display library. It isolated of 104 ELISA-positive clones and sequence analysis showed that 30 clones had full-length light and heavy chains. Four unique gene families were identified, i.e., IgHV3-LV6 (86.66%), IgHV1-LV3 (3.33%), IgHV5-KV3 (3.33%), and IgHV3-LV3 (6.66%). Randomly, one representative clone from each gene family was selected for further studies, i.e., (a) rMAb5 representing IgHV1-LV3, (b) rMAb6 representing IgHV3-LV6, (c) rMAb14 representing IgHV5-KV3, and (d) rMAb23 representing IgHV3-LV3. The rMAb gene sequences from the phage display vector were subcloned into the pET51b+ expression vector and transformed into Escherichia coli Shuffle T7 Express host cell.
format	Thesis
qualification_name	Doctor of Philosophy (PhD.)
qualification_level	Doctorate
author	Balachandra, Dinesh
author_facet	Balachandra, Dinesh
author_sort	Balachandra, Dinesh
title	Isolation, Characterization, And Production Of Recombinant Monoclonal Antibodies Against Rnie For Development Of A Strongyloides Antigen Detection Assay
title_short	Isolation, Characterization, And Production Of Recombinant Monoclonal Antibodies Against Rnie For Development Of A Strongyloides Antigen Detection Assay
title_full	Isolation, Characterization, And Production Of Recombinant Monoclonal Antibodies Against Rnie For Development Of A Strongyloides Antigen Detection Assay
title_fullStr	Isolation, Characterization, And Production Of Recombinant Monoclonal Antibodies Against Rnie For Development Of A Strongyloides Antigen Detection Assay
title_full_unstemmed	Isolation, Characterization, And Production Of Recombinant Monoclonal Antibodies Against Rnie For Development Of A Strongyloides Antigen Detection Assay
title_sort	isolation, characterization, and production of recombinant monoclonal antibodies against rnie for development of a strongyloides antigen detection assay
granting_institution	Universiti Sains Malaysia
granting_department	Institut Penyelidikan Perubatan Molekul
publishDate	2022
url	http://eprints.usm.my/59388/1/DINESH%20AL%20BALACHANDRA%20-%20TESIS24.pdf
_version_	1783729546163388416

Isolation, Characterization, And Production Of Recombinant Monoclonal Antibodies Against Rnie For Development Of A Strongyloides Antigen Detection Assay

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