Murine myeloma cell line

Immunization and Fusion Protocol. Groups of five BALB/c female mice (4-6 weeks old), were given series of three injections every two weeks, with KLH-conjugated atrazine or hydroxyatrazine (50 ug/injection) mixed with Freund s adjuvant. After a rest period of two months, the mice were boosted with 500 ug of the conjugate. Three to four days later, the mice were sacrificed and the spleen cells were fused with the murine myeloma cell line Sp 2/0.Agl4 (12,13). The positive hybridomas were cloned and expanded in mice and the MAbs were purified from the ascitic fluid by ammonium sulfate precipitation, and DEAE-cellulose anion-exchange chromatography (14). 

The next most commonly used cell lines are the murine myeloma cell lines such as NS(T and Sp2/0. The NSO cell line is deficient for the glutamine synthetase gene, and the presence of glutamine synthetase (GS) gene on vectors has been used for the selection of stably integrated expression cassettes. ... 

Several s mtheses of nucleotidephospholipids have been reported recently. For example, phospholipid-araC conjugates have been prepared and tested as prodrugs of arotC as inhibitors of the growth of a murine myeloma cell line. The most effective inhibitor was [13, = 1, R = Me(CH2)i4] solubility difficulties may have affected the testing of other analogues. 

Palivizumab Humanized anti-RSV antibody Synagis Antiviral. Humanized mAb (5% murine) directed to an epitope in the A antigenic site of the F protein of respiratory syncytial virus (RSV). Produced by a stable murine myeloma cell line. respiratory diseases caused by respiratory syncytial virus... 

Cetuximab (Ebritux ) is used in the treatment of advanced colorectal cancer. It is a chimaeric antibody produced in a murine myeloma cell line. 

Fusion of human lymphocytes with human lymphoblastoid cell lines is a very inefficient process. Fusion of human lymphocytes with murine myeloma cells lead to very unstable hybrids. Upon fusion, preferential loss of human genetic elements is often observed. Unfortunately, particularly common is the loss of chromosomes 2,14 and 22, which encode antibody light and heavy chain loci. The production yields of human monoclonals upon immortalization of the human B-lymphocyte (by whatever means) are also low. 

Immunomodulatory, Immunosupressant. Blocks IL-2R alpha on surface of activated T lymphocytes. Produced by mouse myeloma cell line. Contains human heavy- and light-chain constant regions and murine heavy- and light-chain variable regions. 

Figure 13.5 Outline of the production strategy of CEA-SCAN. The antibody-producing hybridoma cell line was originally obtained by standard methods of hybridoma generation. Spleen-derived murine B-lymphocytes were fused with murine myeloma calls. The resulting stable hybridomas were screened for the production of anti-CEA monoclonals. The clone chosen produces an IgG anti-CEA antibody. Note that the finished product outlined above is not radiolabelled. The freeze-dried antibody preparation (which has a shelf life of 2 years at 2-8 °C) is reconstituted immediately prior to its medical use. The reconstituting solution contains 99mTc, and is formulated to facilitate direct conjugation of the radiolabel to the antibody fragment...

In 1975, Kohler and Milstein reported (1) that immortal cell lines secreting antibody of a single specificity could be produced by the artificial fusion of splenocytes derived from an immune mouse and tumour cells derived from a murine myeloma. They called these cell lines hybridomas and the product from them monoclonal antibodies. The development of monoclonal antibodies opened up huge possibilities in all areas of antibody use because reagents could be created with specificity to a single domain (epitope) on the target... 

Although numerous cell lines have been screened for their efficiency as a host system for recombinant protein production, only a few have shown favorable properties for the expression of biopharmaceuticals (Hauser, 1997). Regulatory and economic issues for large-scale production and the intended application of the recombinant protein (diagnosis, therapy, etc.) have to be carefully considered (Makrides and Prentice, 2003). Three mammalian cell lines are now commonly used by the pharmaceutical industry Chinese hamster ovary (CHO) cells, the murine myeloma SP2/0 and the NS0 cell line (see Table 3.1). These cell lines have been used to produce 11 of 21 therapeutic products approved from 1996 to 2000 (Chu and Robinson, 2001). 

The Chinese hamster ovary (CHO) cell line is now being used as a standard host cell for transfection with genes of interest for later use in the production of recombinant antibodies (Butler, 2005). Other cells with equivalent performance are the murine myelomas NSO and Sp2/0, as well as baby hamster kidney (BHK), human embryonic kidney (HEK-93), and a derivative of the human retina (PER.C6 j (Chu and Robinson, 2001). 

In 1973, Cotton et al. successfully fused cells of two plasmacytoma lines to produce hybrid cells capable of synthesizing both myeloma proteins. Subsequently, hybrid cells derived by fusion of a murine myeloma with spleen cells from appropriately immunized donors were shown to se-... 

Most of the information on DNA rearrangements in switched cells is derived from analysis of B cells that became transformed after switching. The transformation had immortalized the switched phenotype. With few exceptions, switching within transformed cell lines of the B lineage is a rare event. It has been observed in murine pre-B cell lines [27-29], human pre-B and B lymphomas [30], the murine B cell lymphoma 1.29 [31] and in several murine myeloma and hybridoma cell lines (e.g. [32]). In general, the frequencies of switching in these lines do not reflect the switch frequencies in the corresponding B cells from which they are presumed to be derived. 

Numerous cell lines have been developed for the purpose of recombinant protein expression however, several cell lines have dominated the field. Chinese hamster ovary (CHO), NSO and SP2/0 murine myeloma, human embryonic kidney (HEK293), and COS cell fines are popular choices. HeLa, BHK, YB2/0, and PerC6 are several other available options that are not discussed in detail here. Several of these cell fines can be grown in attachment-dependent monolayers or suspension cultures. For large-scale culture, cell suspension is preferred. 

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