Development of a Low Serum Medium for the Production of Monoclonal Antibody Against Congenital Adrenal Hyperplasia by Hybridoma Culture

Development of a Low Serum Medium for the Production of Monoclonal Antibody Against Congenital Adrenal Hyperplasia by Hybridoma Culture

Statistically designed experiments were used in developing a low serum medium for the production of a diagnostic monoclonal antibody against congenital adrenal hyperplasia using Hybridoma 192. A two-level half-fractional factorial design was used for screening six components (Minimum Essential Medi...

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Main Author: Chua, Gek Kee
Format: Article
Language:English
English
Published: Taylor & Francis Group 2016
Subjects:
QR180 Immunology
RM Therapeutics. Pharmacology
TP Chemical technology
Online Access:http://umpir.ump.edu.my/id/eprint/11969/
http://umpir.ump.edu.my/id/eprint/11969/
http://umpir.ump.edu.my/id/eprint/11969/
http://umpir.ump.edu.my/id/eprint/11969/1/Low%20serum%20medium.pdf
http://umpir.ump.edu.my/id/eprint/11969/7/Development%20of%20a%20Low%20Serum%20Medium%20For%20The%20Production%20of%20Monoclonal%20Antibody%20Against%20Congenital%20Adrenal%20Hyperplasia%20by%20Hybridoma%20Culture.pdf
id ump-11969
recordtype eprints
spelling ump-119692017-12-15T04:00:32Z http://umpir.ump.edu.my/id/eprint/11969/ Development of a Low Serum Medium for the Production of Monoclonal Antibody Against Congenital Adrenal Hyperplasia by Hybridoma Culture Chua, Gek Kee QR180 Immunology RM Therapeutics. Pharmacology TP Chemical technology Statistically designed experiments were used in developing a low serum medium for the production of a diagnostic monoclonal antibody against congenital adrenal hyperplasia using Hybridoma 192. A two-level half-fractional factorial design was used for screening six components (Minimum Essential Medium Eagle amino acids, 2-mercaptoethanol, ethanolamine, ferric citrate, zinc sulfate and sodium selenite). The experimental design was then augmented to central composite design. The basal Dulbecco’s modified Eagle’s medium (containing 4 mM L-glutamine, 1% antibiotic- antimycotic agent) supplemented with 0.4% by volume fetal bovine serum, 311.8 mM ferric citrate, 17.3 nM sodium selenite and 4.5 mM zinc sulfate (LSD), was found to support the growth of the hybridoma. Specific cell growth rate in the LSD (0.033 ± 0.001/h) was slightly lower than in the control medium (i.e. basal DMEM supplemented with 2% FBS; 0.0045 ± 0.003/h). Nevertheless, the specific MAb production rate for LSD was higher(0.057 ± 0.015 pg/cell -h versus 0.004 ± 0.002 pg/cell -h in LSD and control respectively). The antibody produced in the LSD showed high specificity and no cross-reactivity with the other structural resemblance’s steroid hormones, revealing no structural changes owing to the new medium developed. The new medium formulation effectively reduced the medium cost by up to 64.6%. Taylor & Francis Group 2016-01 Article PeerReviewed application/pdf en http://umpir.ump.edu.my/id/eprint/11969/1/Low%20serum%20medium.pdf application/pdf en http://umpir.ump.edu.my/id/eprint/11969/7/Development%20of%20a%20Low%20Serum%20Medium%20For%20The%20Production%20of%20Monoclonal%20Antibody%20Against%20Congenital%20Adrenal%20Hyperplasia%20by%20Hybridoma%20Culture.pdf Chua, Gek Kee (2016) Development of a Low Serum Medium for the Production of Monoclonal Antibody Against Congenital Adrenal Hyperplasia by Hybridoma Culture. Preparative Biochemistry and Biotechnology, 46 (7). pp. 679-685. ISSN 1082-6068 http://dx.doi.org/10.1080/10826068.2015.1135450 DOI: 10.1080/10826068.2015.1135450
repository_type Digital Repository
institution_category Local University
institution Universiti Malaysia Pahang
building UMP Institutional Repository
collection Online Access
language English
English
topic QR180 Immunology
RM Therapeutics. Pharmacology
TP Chemical technology
spellingShingle QR180 Immunology
RM Therapeutics. Pharmacology
TP Chemical technology
Chua, Gek Kee
Development of a Low Serum Medium for the Production of Monoclonal Antibody Against Congenital Adrenal Hyperplasia by Hybridoma Culture
description Statistically designed experiments were used in developing a low serum medium for the production of a diagnostic monoclonal antibody against congenital adrenal hyperplasia using Hybridoma 192. A two-level half-fractional factorial design was used for screening six components (Minimum Essential Medium Eagle amino acids, 2-mercaptoethanol, ethanolamine, ferric citrate, zinc sulfate and sodium selenite). The experimental design was then augmented to central composite design. The basal Dulbecco’s modified Eagle’s medium (containing 4 mM L-glutamine, 1% antibiotic- antimycotic agent) supplemented with 0.4% by volume fetal bovine serum, 311.8 mM ferric citrate, 17.3 nM sodium selenite and 4.5 mM zinc sulfate (LSD), was found to support the growth of the hybridoma. Specific cell growth rate in the LSD (0.033 ± 0.001/h) was slightly lower than in the control medium (i.e. basal DMEM supplemented with 2% FBS; 0.0045 ± 0.003/h). Nevertheless, the specific MAb production rate for LSD was higher(0.057 ± 0.015 pg/cell -h versus 0.004 ± 0.002 pg/cell -h in LSD and control respectively). The antibody produced in the LSD showed high specificity and no cross-reactivity with the other structural resemblance’s steroid hormones, revealing no structural changes owing to the new medium developed. The new medium formulation effectively reduced the medium cost by up to 64.6%.
format Article
author Chua, Gek Kee
author_facet Chua, Gek Kee
author_sort Chua, Gek Kee
title Development of a Low Serum Medium for the Production of Monoclonal Antibody Against Congenital Adrenal Hyperplasia by Hybridoma Culture
title_short Development of a Low Serum Medium for the Production of Monoclonal Antibody Against Congenital Adrenal Hyperplasia by Hybridoma Culture
title_full Development of a Low Serum Medium for the Production of Monoclonal Antibody Against Congenital Adrenal Hyperplasia by Hybridoma Culture
title_fullStr Development of a Low Serum Medium for the Production of Monoclonal Antibody Against Congenital Adrenal Hyperplasia by Hybridoma Culture
title_full_unstemmed Development of a Low Serum Medium for the Production of Monoclonal Antibody Against Congenital Adrenal Hyperplasia by Hybridoma Culture
title_sort development of a low serum medium for the production of monoclonal antibody against congenital adrenal hyperplasia by hybridoma culture
publisher Taylor & Francis Group
publishDate 2016
url http://umpir.ump.edu.my/id/eprint/11969/
http://umpir.ump.edu.my/id/eprint/11969/
http://umpir.ump.edu.my/id/eprint/11969/
http://umpir.ump.edu.my/id/eprint/11969/1/Low%20serum%20medium.pdf
http://umpir.ump.edu.my/id/eprint/11969/7/Development%20of%20a%20Low%20Serum%20Medium%20For%20The%20Production%20of%20Monoclonal%20Antibody%20Against%20Congenital%20Adrenal%20Hyperplasia%20by%20Hybridoma%20Culture.pdf
first_indexed 2023-09-18T22:13:05Z
last_indexed 2023-09-18T22:13:05Z
_version_ 1777415157913223168

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