Effects of culture conditions on the production and quality of monoclonal IgA

Stoll, T. S.; Chappaz, A.; Stockar, Urs von; Marison, I. W.

doi:10.1016/s0141-0229(96)00268-2

Cited by 10 publications

(4 citation statements)

References 20 publications

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“…This Western blot clearly demonstrated that cells fused to produce all the four polypeptides of SIgA, assembled and secreted SIgA of the expected molecular size with reactivity against antibodies towards both IgA and SC. The fused cells expressed both monomeric and polymeric IgA as described for hybridoma cells expressing monoclonal IgA [27], and was also seen when the J‐chain gene was transfected into an IgA‐producing CHO cell line [2]. The production level of the fused cells grown in culture, without any selective pressure, was measured by ELISA for two months.…”

Section: Resultsmentioning

confidence: 99%

Balanced expression of single subunits in a multisubunit protein, achieved by cell fusion of individual transfectants

Norderhaug¹,

Johansen

Sandlie

2002

European Journal of Biochemistry

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To establish stable cell lines that produce recombinant multisubunit proteins, it is usually necessary to cotransfect cells with several independent gene constructs. Here, we show that a stepwise fusion of individually transfected cells, results in a fused cell-line that secretes a complete multisubunit protein. Functional expression of recombinant multisubunit proteins may require a defined expression ratio between each protein subunit. The cell-fusion technology described allows a predefined expression level of each subunit. Using SIgA as a model protein we demonstrate that the majority of the fused cells inherit the molar expression ratio of the parental transfected cells. These results indicate that simplified screening of clones expressing the expected subunit ratios may be possible using the cell-fusion technology. This technology may therefore be an alternative to generic transfection methods for the establishment of cells that produce multiprotein complexes such as antibodies, receptors, ion channels and other multisubunit proteins.

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Section: Resultsmentioning

confidence: 99%

Balanced expression of single subunits in a multisubunit protein, achieved by cell fusion of individual transfectants

Norderhaug¹,

Johansen

Sandlie

2002

European Journal of Biochemistry

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“…Several kinds of animal cell bioreactors are presently being used both for industrial production and in pilot laboratory research. For example, stirred tank reactor,12, 13 airlift bioreactor, radial flow,14 fixed‐bed,15, 16 fibrous bed,17, 18 rolling culture system,19 coiled tube membrane,20 rotary cell culture system,21 wave bioreactor culture system,22, 23 and hollow fiber24–26 have been reported. Anchorage‐dependent animal cells need to attach themselves to a solid surface to grow, and if the solid surface in a culture vessel is increased, the final cell density will increase correspondingly.…”

Section: Introductionmentioning

confidence: 99%

Three‐dimensional culture for monoclonal antibody production by hybridoma cells immobilized in macroporous gel particles

Nilsang

Nehru

Plieva

et al. 2008

Biotechnology Progress

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in Wiley InterScience (www.interscience.wiley.com).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 Â 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 þ 2 mM a-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 þ a-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 Â 10 6 cells mL À1) and highest specific mAb production rate (3.9 lg mL À1 10 À4 viable cell day À1), 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.

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“…However, these devices can only produce 1−2 L of culture per batch due to their inherently limited oxygen‐transfer capabilities. For larger quantities, there are many different types of cell culture bioreactor systems available, for example, stirred tank (5, 6), radial flow (7), fixed‐bed (8, 9), fibrous‐bed (10, 11), hollow fiber (12 − 14), and wave bioreactor (15, 16). High cell density and good long‐term culture stability are two key factors in developing a continuous bioreactor system.…”

Section: Introductionmentioning

confidence: 99%

Monoclonal Antibody Production Using a New Supermacroporous Cryogel Bioreactor

Nilsang¹,

Nandakumar²,

Galaev³

et al. 2007

Biotechnol. Prog.

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A supermacroporous cryogel bioreactor has been developed to culture hybridoma cells for long-term continuous production of monoclonal antibodies (mAb). Hybridoma clone M2139, secreting antibodies against J1 epitope (GERGAAGIAGPK; amino acids, 551-564) of collagen type II, are immobilized in the porous bed matrix of a cryogel column (10 mL bed volume). The cells got attached to the matrix within 48 h after inoculation and grew as a confluent sheet inside the cryogel matrix. Cells were in the lag phase for 15 days and secreted mAb into the circulation medium. Glucose consumption and lactic acid production were also monitored, and during the exponential phase (approximately 20 days), the hybridoma cell line consumed 0.75 mM day-1 glucose, produced 2.48 mM day-1 lactic acid, and produced 6.5 microg mL-1 day-1 mAb during the exponential phase. The mAb concentration reached 130 microg mL-1 after continuous run of the cryogel column for 36 days. The yield of the mAb after purification was 67.5 mg L-1, which was three times greater than the mAb yield obtained from T-flask batch cultivation. Even after the exchange of medium reservoir, cells in the cryogel column were still active and had relatively stable mAb production for an extended period of time. The bioreactor was operated continuously for 55 days without any contamination. The results from ELISA as well as arthritis experiments demonstrate that the antibodies secreted by cells grown on the cryogel column did not differ from antibodies purified from the cells grown in commercial CL-1000 culture flasks. Thus, supermacroporous cryogels can be useful as a supporting material for productive hybridoma cell culture. Cells were found to be viable inside the porous matrix of the cryogel during the study period and secreted antibodies continuously. The antibodies thus obtained from the cryogel reactor were found to be functionally active in vivo, as demonstrated by their capacity to induce arthritis in mice.

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Effects of culture conditions on the production and quality of monoclonal IgA

Cited by 10 publications

References 20 publications

Balanced expression of single subunits in a multisubunit protein, achieved by cell fusion of individual transfectants

Balanced expression of single subunits in a multisubunit protein, achieved by cell fusion of individual transfectants

Three‐dimensional culture for monoclonal antibody production by hybridoma cells immobilized in macroporous gel particles

Monoclonal Antibody Production Using a New Supermacroporous Cryogel Bioreactor

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