Effect of α-Ketoglutarate on Monoclonal Antibody Production of Hybridoma Cell Lines in Serum-Free and Serum-Containing Medium

Nilsang, Suthasinee; Kumar, Ashok; Rakshit, Sudip Kumar

doi:10.1007/s12010-008-8225-0

Cited by 16 publications

(12 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…These results revealed that supplementation of 2 mM α‐ketoglutarate in the normal glutamine‐containing D‐MEM medium can enhance cell growth and mAb production. Similar results have also been observed with other hybridoma cell lines in other serum containing and serum‐free medium 50. However, this experiment may further need to be optimized for specific amount of α‐ketoglutarate supplement in the media and also some other parameters such as ammonia accumulation need to be checked.…”

Section: Resultssupporting

confidence: 83%

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

Nilsang

Nehru

Plieva

et al. 2008

Biotechnology Progress

Self Cite

View full text Add to dashboard Cite

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.

show abstract

Section: Resultssupporting

confidence: 83%

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

Nilsang

Nehru

Plieva

et al. 2008

Biotechnology Progress

Self Cite

View full text Add to dashboard Cite

show abstract

“…In this study it was clear that enhanced cell proliferation could be achieved by adding α-KG in the culture medium. In our previous study improved production of monoclonal antibody was achieved by the addition of α-KG in medium 2. Macroporous cryogel matrix helped in the better nutrient transport and cell growth and addition of α-KG prevented the accumulation of the toxic catabolites resulting in improved culture stability.…”

Section: Resultsmentioning

confidence: 99%

“…One of the most important components for production of these biological products is primary or immortal cells. The cells cultured in laboratory conditions has specific requirements in which the cell media plays an important role 2 but frequent accumulation of toxic catabolic components like ammonia effects the overall cellular growth which directly effects the therapeutic production 3.…”

Section: Introductionmentioning

confidence: 99%

Effect of Alpha-Ketoglutarate on Growth and Metabolism of Cells Cultured on Three-Dimensional Cryogel Matrix

Singh

Vishnoi²,

Kumar

2013

Int. J. Biol. Sci.

Self Cite

View full text Add to dashboard Cite

Alpha- ketoglutarate (α-KG) is a well-known intermediate of Kreb's cycle and thus one of the important candidates in the role of cellular metabolism. The aim was to investigate the effect of α-KG on the growth and proliferation of different cells (fibroblasts and chondrocytes). Further change in the consumption of glucose and release of ammonia in the media, upon the addition of α-KG was also examined. NIH3T3 and chondrocytes were seeded on two different macroporous cryogel matrices synthesized by poly (hydroxyethylmethaacrylate)-gelatin (HG) and alginate-gelatin (AG). Higher proliferation was observed for both the cell lines in 2-D as well as 3-D where α-KG was added to the media compared to the controls. It was evident that the chondrocytes were metabolically more active in the case of α-KG containing samples as early extracellular matrix (ECM) accumulation was observed. In comparison to the 6-8 weeks duration required by the chondrocytes for ECM accumulation in normal in-vitro culture conditions, α-KG containing samples showed an earlier accumulation within 3 weeks. In order to further validate the results, scanning electron microscopic (SEM) analysis was performed showed high ECM deposition and cells embedded in the matrix. Homogenous distribution of cells on both the synthesized matrix was reported using 4'-6-diamidino-2-phenylindole (DAPI) and propidium iodide (PI) staining. The results clearly showed reduction in ammonia concentration in α-KG containing samples thus leading to decreased ammonia toxicity that builds up during long cell culture conditions thereby enhancing the proliferation and metabolic activity. Thus α-KG can be used potentially for long batch-cultures for the production of vaccines or antibody as well as can play an important role as a bioactive molecule for in vitro neo-cartilage generation.

show abstract

“…The modified high affinity xfR5 mAb were produced and isolated from a Hybridoma cell line, i.e., XF27/28/CCR5 43E2-AA (PTA-4054; ATCC repository) (54), by following published method with modifications as described below (56). Briefly, XF-CCR5 28/27 43E2AA hybridoma cells were seeded (at 10 6 /mL concentration) and maintained in antibody production inducing media, i.e., RPMI medium with 1×AA, for several days until 50% cells were found to be compromised (cell death).…”

Section: Methodsmentioning

confidence: 99%

Targeted immuno-antiretroviral HIV therapeutic approach to provide dual protection and boosts cellular immunity: A proof-of-concept study

Mandal¹,

Prathipati²,

Belshan³

et al. 2020

Preprint

View full text Add to dashboard Cite

Human immunodeficiency virus (HIV)-infected active and latent CCR5 expressing long-lived T-cells are the primary barrier to HIV/AIDS eradication. Broadly neutralizing antibodies and latency-reversing agents are the two most promising strategies emerging to achieve ‘functional cure’ against HIV infection. Antiretrovirals (ARVs) have shown to suppress plasma viral loads to non-detectable levels and above strategies have demonstrated a ‘functional cure’ against HIV infection is achievable. Both the above strategies are effective at inducing direct or immune-mediated cell death of latent HIV+ T-cells but have shown respective limitations. In this study, we designed a novel targeted ARVs-loaded nanoformulation that combines the CCR5 monoclonal antibody and antiretroviral drugs (ARV) as a dual protection strategy to promote HIV ‘functional cure’. The modified CCR5 monoclonal antibody (xfR5 mAb) surface-coated dolutegravir (DTG) and tenofovir alafenamide (TAF) loaded nanoformulation (xfR5-D+T NPs) were uniformly sized <250 nm, with 6.5 times enhanced antigen-binding affinity compared to naïve xfR5 mAb, and provided prolonged DTG and TAF intracellular retention (t1/2). The multivalent and sustained drug release properties of xfR5-D+T NPs enhance the protection efficiency against HIV by approximately 12, 3, and 5 times compared to naïve xfR5 mAb, D+T NP alone, and xfR5 NPs, respectively. Further, the nanoformulation demonstrated high binding-affinity to CCR5 expressing CD4+ cells, monocytes, and other HIV prone/latent T-cells by 25, 2, and 2 times, respectively. Further, the xfR5-D+T NPs during short-term pre-exposure prophylaxis induced a protective immunophenotype, i.e., boosted T-helper (Th), temporary memory (TM), and effector (E) sub-population. Moreover, treatment with xfR5-D+T NPs to HIV-infected T-cells induced a defensive/activated immunophenotype i.e., boosted naïve, Th, boosted central memory, TM, EM, E, and activated cytotoxic T-cells population. Therefore, this dual-action targeted mAb-ARV loaded nanoformulation could potentially become a multifactorial/multilayered solution to achieve a “functional cure.”

show abstract

Effect of α-Ketoglutarate on Monoclonal Antibody Production of Hybridoma Cell Lines in Serum-Free and Serum-Containing Medium

Cited by 16 publications

References 25 publications

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

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

Effect of Alpha-Ketoglutarate on Growth and Metabolism of Cells Cultured on Three-Dimensional Cryogel Matrix

Targeted immuno-antiretroviral HIV therapeutic approach to provide dual protection and boosts cellular immunity: A proof-of-concept study

Contact Info

Product

Resources

About