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Three-Dimensional Culture for Monoclonal Antibody Production by Hybridoma Cells Immobilized in Macroporous Gel Particles

Nilsang, Suthasinee LU ; Nehru, Vishal LU ; Plieva, Fatima LU ; Kutty Selva, Nandakumar LU ; Rakshit, Sudip Kumar; Holmdahl, Rikard LU ; Mattiasson, Bo LU and Kumar, Ashok LU (2008) In Biotechnology Progress 24(5). p.1122-1131
Abstract
Cell proliferation and long-term production of monoclonal antibody IgG(2b) by M2139 hybridoma cells immobilized in macroporous gel particles (MGPs) in packed-bed reactor were studied for a period of 60 days. The MGPs were made of supermacroporous gels produced in frozen conditions from crosslinked polyacrylamide and modified with gelatin which were housed in special plastic carriers (7 x 9 mm(2)). Cells were trapped in the interior part of MGPs by attaching to the void space of the gel matrix as three-dimensional (3D) cultivation using gelatin as a substrate layer. Optimizing productivity by hybridoma cell relies on understanding regulation of antibody production. In this study, the behavior of M2139 cells in two-dimensional cultures on... (More)
Cell proliferation and long-term production of monoclonal antibody IgG(2b) by M2139 hybridoma cells immobilized in macroporous gel particles (MGPs) in packed-bed reactor were studied for a period of 60 days. The MGPs were made of supermacroporous gels produced in frozen conditions from crosslinked polyacrylamide and modified with gelatin which were housed in special plastic carriers (7 x 9 mm(2)). Cells were trapped in the interior part of MGPs by attaching to the void space of the gel matrix as three-dimensional (3D) cultivation using gelatin as a substrate layer. Optimizing productivity by hybridoma cell relies on understanding regulation of antibody production. In this study, the behavior of M2139 cells in two-dimensional cultures on multiwell plate surfaces was also investigated. The effect of three different medium such as basal medium Dulbecco's modified Eagle's medium (D-MEM) containing L-glutamine or L-glutamine + 2 mM alpha-ketoglutarate or L-alanyl-glutamine (GlutaMAX (TM) ) was studied prior to its use in 3D cultivation. The kinetics of cell growth in basal medium containing L-glutamine + alpha-ketoglutarate was similar to cells grown on Gluta- MAX containing medium, whereas D-MEM containing L-glutamine showed lower productivity. With the maximal viable cell density (6.85 x 10(6) cells mL(-1)) and highest specific mAb production rate (3.9 mu g mL(-1) 10(-4) viable cell day(-1)), D-MEM-GlutaMAX was further selected for 3D cultivation. Cells in MGPs were able to grow and secrete antibody for 30 days in packed-bed batch reactor, before a fresh medium reservoir was replaced. After being supplied with fresh medium, cells again showed continuous growth for another 30 days with mAb production efficiency of 50%. These results demonstrate that MGPs can be used efficiently as supporting carrier for long-term monoclonal antibody production. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
macroporous gel particles, antibody production, M2139 hybridoma cells, 3D cultivation, medium optimization, cell immobilization
in
Biotechnology Progress
volume
24
issue
5
pages
1122 - 1131
publisher
The American Chemical Society
external identifiers
  • wos:000260738900015
  • pmid:19194922
  • scopus:56249132172
ISSN
1520-6033
DOI
10.1002/btpr.28
language
English
LU publication?
yes
id
5f44a9ac-e6df-494c-8d7f-84601d214f11 (old id 1283665)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/19194922?dopt=Abstract
date added to LUP
2009-02-10 10:23:31
date last changed
2017-07-30 03:53:53
@article{5f44a9ac-e6df-494c-8d7f-84601d214f11,
  abstract     = {Cell proliferation and long-term production of monoclonal antibody IgG(2b) by M2139 hybridoma cells immobilized in macroporous gel particles (MGPs) in packed-bed reactor were studied for a period of 60 days. The MGPs were made of supermacroporous gels produced in frozen conditions from crosslinked polyacrylamide and modified with gelatin which were housed in special plastic carriers (7 x 9 mm(2)). Cells were trapped in the interior part of MGPs by attaching to the void space of the gel matrix as three-dimensional (3D) cultivation using gelatin as a substrate layer. Optimizing productivity by hybridoma cell relies on understanding regulation of antibody production. In this study, the behavior of M2139 cells in two-dimensional cultures on multiwell plate surfaces was also investigated. The effect of three different medium such as basal medium Dulbecco's modified Eagle's medium (D-MEM) containing L-glutamine or L-glutamine + 2 mM alpha-ketoglutarate or L-alanyl-glutamine (GlutaMAX (TM) ) was studied prior to its use in 3D cultivation. The kinetics of cell growth in basal medium containing L-glutamine + alpha-ketoglutarate was similar to cells grown on Gluta- MAX containing medium, whereas D-MEM containing L-glutamine showed lower productivity. With the maximal viable cell density (6.85 x 10(6) cells mL(-1)) and highest specific mAb production rate (3.9 mu g mL(-1) 10(-4) viable cell day(-1)), D-MEM-GlutaMAX was further selected for 3D cultivation. Cells in MGPs were able to grow and secrete antibody for 30 days in packed-bed batch reactor, before a fresh medium reservoir was replaced. After being supplied with fresh medium, cells again showed continuous growth for another 30 days with mAb production efficiency of 50%. These results demonstrate that MGPs can be used efficiently as supporting carrier for long-term monoclonal antibody production.},
  author       = {Nilsang, Suthasinee and Nehru, Vishal and Plieva, Fatima and Kutty Selva, Nandakumar and Rakshit, Sudip Kumar and Holmdahl, Rikard and Mattiasson, Bo and Kumar, Ashok},
  issn         = {1520-6033},
  keyword      = {macroporous gel particles,antibody production,M2139 hybridoma cells,3D cultivation,medium optimization,cell immobilization},
  language     = {eng},
  number       = {5},
  pages        = {1122--1131},
  publisher    = {The American Chemical Society},
  series       = {Biotechnology Progress},
  title        = {Three-Dimensional Culture for Monoclonal Antibody Production by Hybridoma Cells Immobilized in Macroporous Gel Particles},
  url          = {http://dx.doi.org/10.1002/btpr.28},
  volume       = {24},
  year         = {2008},
}