Evaluation of multi-well based system for antibody production using Chinese hamster ovary cell

Evaluation of multi-well based system for antibody production using Chinese hamster ovary cell

The increased use of antibodies and antibody fragments for human therapy is driving the search for rational approaches to accelerate bioprocess development. However, screening of the operating conditions of process parameters in large scale bioreactors for biopharmaceuticals industry is not cost eff...

Full description

Saved in:
Bibliographic Details
Main Author: Sabri, Fatin Syamimi
Format: Thesis
Language:English
Published: 2018
Subjects:
QH301 Biology
Online Access:http://eprints.utm.my/id/eprint/85828/1/FatinSyamimiSabriMFBME2018.pdf
Tags: Add Tag
No Tags, Be the first to tag this record!
id my-utm-ep.85828
record_format uketd_dc
spelling my-utm-ep.858282020-07-30T07:35:06Z Evaluation of multi-well based system for antibody production using Chinese hamster ovary cell 2018 Sabri, Fatin Syamimi QH301 Biology The increased use of antibodies and antibody fragments for human therapy is driving the search for rational approaches to accelerate bioprocess development. However, screening of the operating conditions of process parameters in large scale bioreactors for biopharmaceuticals industry is not cost effective. Using microwell plate as the 'microreactor' for screening process wrould be a solution to this. Hence, this study was carried out to evaluate multi-well based system by determining the growth rate and antibody expression of the Chinese hamster ovary (CHO) cells. In this study, two types of CHO cells have been seeded with different concentrations which were 7^10^ cell/ml and lxlO6 cell/ml. Wild type CHO cell and CHO DG44 were used in this study. The wild type CHO cell was used as a reference cell as it is a stable cell. The cell number wras calculated by using trypan blue exclusion method while glucose concentration was determined by 3. 5- Dinitrosalicyclic acid (DNS) assay. On the other hand, antibody production was determined by using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. For the cell growth rate and doubling time, CHO DG44 (lxlO6 cell/ml) has recorded the highest growth rate and shortest doubling time with (0.3301 ± 0.1536)/h and (2.61 ±1.66)h. Besides that the highest glucose concentration recorded by wild type CHO cell (lxlO6 cell/ml) and CHO DG44 (7*10^ cell/ml) was on day 0 of incubation with (1,817±0.1187)mg/ml and (1.819±0.9331)mg/ml. Meanwhile, CHO DG44 (1 * 106 cell/ml) has recorded the highest glucose concentration on day 1 with (1.61 l±0.1273)mg/ml. Lastly, the antibodyexpress ion by CHO DG44 was verified. Both batches of CHO DG44 have successfully expressed human IgG on day 2 until day 10. In conclusion, the CHO cell is possible to be cultured in the multi-well plate. 2018 Thesis http://eprints.utm.my/id/eprint/85828/ http://eprints.utm.my/id/eprint/85828/1/FatinSyamimiSabriMFBME2018.pdf application/pdf en public http://dms.library.utm.my:8080/vital/access/manager/Repository/vital:132658 masters Universiti Teknologi Malaysia, Faculty of Engineering - School of Biomedical Engineering & Health Sciences Faculty of Engineering - School of Biomedical Engineering & Health Sciences
institution Universiti Teknologi Malaysia
collection UTM Institutional Repository
language English
topic QH301 Biology
spellingShingle QH301 Biology
Sabri, Fatin Syamimi
Evaluation of multi-well based system for antibody production using Chinese hamster ovary cell
description The increased use of antibodies and antibody fragments for human therapy is driving the search for rational approaches to accelerate bioprocess development. However, screening of the operating conditions of process parameters in large scale bioreactors for biopharmaceuticals industry is not cost effective. Using microwell plate as the 'microreactor' for screening process wrould be a solution to this. Hence, this study was carried out to evaluate multi-well based system by determining the growth rate and antibody expression of the Chinese hamster ovary (CHO) cells. In this study, two types of CHO cells have been seeded with different concentrations which were 7^10^ cell/ml and lxlO6 cell/ml. Wild type CHO cell and CHO DG44 were used in this study. The wild type CHO cell was used as a reference cell as it is a stable cell. The cell number wras calculated by using trypan blue exclusion method while glucose concentration was determined by 3. 5- Dinitrosalicyclic acid (DNS) assay. On the other hand, antibody production was determined by using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. For the cell growth rate and doubling time, CHO DG44 (lxlO6 cell/ml) has recorded the highest growth rate and shortest doubling time with (0.3301 ± 0.1536)/h and (2.61 ±1.66)h. Besides that the highest glucose concentration recorded by wild type CHO cell (lxlO6 cell/ml) and CHO DG44 (7*10^ cell/ml) was on day 0 of incubation with (1,817±0.1187)mg/ml and (1.819±0.9331)mg/ml. Meanwhile, CHO DG44 (1 * 106 cell/ml) has recorded the highest glucose concentration on day 1 with (1.61 l±0.1273)mg/ml. Lastly, the antibodyexpress ion by CHO DG44 was verified. Both batches of CHO DG44 have successfully expressed human IgG on day 2 until day 10. In conclusion, the CHO cell is possible to be cultured in the multi-well plate.
format Thesis
qualification_level Master's degree
author Sabri, Fatin Syamimi
author_facet Sabri, Fatin Syamimi
author_sort Sabri, Fatin Syamimi
title Evaluation of multi-well based system for antibody production using Chinese hamster ovary cell
title_short Evaluation of multi-well based system for antibody production using Chinese hamster ovary cell
title_full Evaluation of multi-well based system for antibody production using Chinese hamster ovary cell
title_fullStr Evaluation of multi-well based system for antibody production using Chinese hamster ovary cell
title_full_unstemmed Evaluation of multi-well based system for antibody production using Chinese hamster ovary cell
title_sort evaluation of multi-well based system for antibody production using chinese hamster ovary cell
granting_institution Universiti Teknologi Malaysia, Faculty of Engineering - School of Biomedical Engineering & Health Sciences
granting_department Faculty of Engineering - School of Biomedical Engineering & Health Sciences
publishDate 2018
url http://eprints.utm.my/id/eprint/85828/1/FatinSyamimiSabriMFBME2018.pdf
_version_ 1747818460100952064

Similar Items