Development of monospecific polyclonal antibodies against hypervirulent Klebsiella pneumoniae

Abstract. Darniati, Setiyaningsih S, Agungpriyono DR, Handharyani E. 2021. Development of monospecific polyclonal antibodies against hypervirulent Klebsiella pneumoniae. Biodiversitas 22: 99-105. The aim of the research was to produce monospecific antibodies against Klebsiella pneumoniae serotype K1...

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Autores principales:	darniati darniati, Surachmi Setiyaningsih, Dewi Ratih Agungpriyono, Ekowati Handharyani
Formato:	article
Lenguaje:	EN
Publicado:	MBI & UNS Solo 2020
Materias:	immunoaffinity purification, klebsiella pneumoniae, monospecific antibody, sds-page Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/3c8b28ad83f8409187f73d8021abdddb
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spelling	oai:doaj.org-article:3c8b28ad83f8409187f73d8021abdddb2021-11-22T00:53:57ZDevelopment of monospecific polyclonal antibodies against hypervirulent Klebsiella pneumoniae1412-033X2085-472210.13057/biodiv/d220114https://doaj.org/article/3c8b28ad83f8409187f73d8021abdddb2020-12-01T00:00:00Zhttps://smujo.id/biodiv/article/view/7224https://doaj.org/toc/1412-033Xhttps://doaj.org/toc/2085-4722Abstract. Darniati, Setiyaningsih S, Agungpriyono DR, Handharyani E. 2021. Development of monospecific polyclonal antibodies against hypervirulent Klebsiella pneumoniae. Biodiversitas 22: 99-105. The aim of the research was to produce monospecific antibodies against Klebsiella pneumoniae serotype K1 and K2. Klebsiella pneumonia isolates were recovered from pneumonic lungs of Aceh cattle. The bacteria were identified by rpoB (RNA polymerase ? subunit) PCR amplification specific for K. pneumoniae. The presence of capsule cluster gene magA and k2A was used to characterize capsular serotype K1 and K2, respectively. Antisera were produced using serial immunization of New Zealand White rabbits with K. pneumoniae serotypes K1 and K2. The presence of antibodies was determined by using immunodiffusion test and purification was performed by immunoaffinity purification method. K. pneumoniae antibodies began to appear on the 14th day after the first injection and progressively intensified after the 1st and 2nd booster. The cross-reactivity between the antigen was eliminated by absorbing the antisera with the opposite antigen and several serotypes of non-K1/K2. After purification, serum protein concentrations were substantially decreased from 58.48 µg/µL and 53.99 µg/µL to 2.38 µg / µL and 3.72 µg / µL for K1 and K2, respectively). The SDS-PAGE analysis showed two bands with molecular weight at 54 kDa and 25 kDa representing the heavy and light chains of immunoglobulin G, respectively. Purified polyclonal antiseras against K. pneumoniae serotypes K1 and K2 showed high affinity and specificity for homologous antigens in immunodiffusion and direct agglutination tests, and immunohistochemical staining.darniati darniatiSurachmi SetiyaningsihDewi Ratih AgungpriyonoEkowati HandharyaniMBI & UNS Soloarticleimmunoaffinity purification, klebsiella pneumoniae, monospecific antibody, sds-pageBiology (General)QH301-705.5ENBiodiversitas, Vol 22, Iss 1 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	immunoaffinity purification, klebsiella pneumoniae, monospecific antibody, sds-page Biology (General) QH301-705.5
spellingShingle	immunoaffinity purification, klebsiella pneumoniae, monospecific antibody, sds-page Biology (General) QH301-705.5 darniati darniati Surachmi Setiyaningsih Dewi Ratih Agungpriyono Ekowati Handharyani Development of monospecific polyclonal antibodies against hypervirulent Klebsiella pneumoniae
description	Abstract. Darniati, Setiyaningsih S, Agungpriyono DR, Handharyani E. 2021. Development of monospecific polyclonal antibodies against hypervirulent Klebsiella pneumoniae. Biodiversitas 22: 99-105. The aim of the research was to produce monospecific antibodies against Klebsiella pneumoniae serotype K1 and K2. Klebsiella pneumonia isolates were recovered from pneumonic lungs of Aceh cattle. The bacteria were identified by rpoB (RNA polymerase ? subunit) PCR amplification specific for K. pneumoniae. The presence of capsule cluster gene magA and k2A was used to characterize capsular serotype K1 and K2, respectively. Antisera were produced using serial immunization of New Zealand White rabbits with K. pneumoniae serotypes K1 and K2. The presence of antibodies was determined by using immunodiffusion test and purification was performed by immunoaffinity purification method. K. pneumoniae antibodies began to appear on the 14th day after the first injection and progressively intensified after the 1st and 2nd booster. The cross-reactivity between the antigen was eliminated by absorbing the antisera with the opposite antigen and several serotypes of non-K1/K2. After purification, serum protein concentrations were substantially decreased from 58.48 µg/µL and 53.99 µg/µL to 2.38 µg / µL and 3.72 µg / µL for K1 and K2, respectively). The SDS-PAGE analysis showed two bands with molecular weight at 54 kDa and 25 kDa representing the heavy and light chains of immunoglobulin G, respectively. Purified polyclonal antiseras against K. pneumoniae serotypes K1 and K2 showed high affinity and specificity for homologous antigens in immunodiffusion and direct agglutination tests, and immunohistochemical staining.
format	article
author	darniati darniati Surachmi Setiyaningsih Dewi Ratih Agungpriyono Ekowati Handharyani
author_facet	darniati darniati Surachmi Setiyaningsih Dewi Ratih Agungpriyono Ekowati Handharyani
author_sort	darniati darniati
title	Development of monospecific polyclonal antibodies against hypervirulent Klebsiella pneumoniae
title_short	Development of monospecific polyclonal antibodies against hypervirulent Klebsiella pneumoniae
title_full	Development of monospecific polyclonal antibodies against hypervirulent Klebsiella pneumoniae
title_fullStr	Development of monospecific polyclonal antibodies against hypervirulent Klebsiella pneumoniae
title_full_unstemmed	Development of monospecific polyclonal antibodies against hypervirulent Klebsiella pneumoniae
title_sort	development of monospecific polyclonal antibodies against hypervirulent klebsiella pneumoniae
publisher	MBI & UNS Solo
publishDate	2020
url	https://doaj.org/article/3c8b28ad83f8409187f73d8021abdddb
work_keys_str_mv	AT darniatidarniati developmentofmonospecificpolyclonalantibodiesagainsthypervirulentklebsiellapneumoniae AT surachmisetiyaningsih developmentofmonospecificpolyclonalantibodiesagainsthypervirulentklebsiellapneumoniae AT dewiratihagungpriyono developmentofmonospecificpolyclonalantibodiesagainsthypervirulentklebsiellapneumoniae AT ekowatihandharyani developmentofmonospecificpolyclonalantibodiesagainsthypervirulentklebsiellapneumoniae
_version_	1718418452800077824

Development of monospecific polyclonal antibodies against hypervirulent Klebsiella pneumoniae

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