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Monoclonal antibodies myeloma cell preparation

Figure 10.3. Schematic representation of monoclonal antibody production using immortalized hybrid cells that secrete antibodies selective for the target antigen. The mortal, immune B cells Isolated from mice immunized with a target antigen are fused with myeloma, immortal B cells that express defective antibodies. The selecting of antigen-specific, immortal hybrid cells (hybridomas) results in identification of unique clones of cells that express antibodies with high specificity and affinity (monoclonal antibodies). These cells are cloned and expanded for large-scale monoclonal antibody preparations. Figure 10.3. Schematic representation of monoclonal antibody production using immortalized hybrid cells that secrete antibodies selective for the target antigen. The mortal, immune B cells Isolated from mice immunized with a target antigen are fused with myeloma, immortal B cells that express defective antibodies. The selecting of antigen-specific, immortal hybrid cells (hybridomas) results in identification of unique clones of cells that express antibodies with high specificity and affinity (monoclonal antibodies). These cells are cloned and expanded for large-scale monoclonal antibody preparations.
Figure 4.33. Preparation of Monoclonal Antibodies. Hybridoma cells are formed by fusion of antibody-producing cells and myeloma cells. The hybrid cells are allowed to proliferate by growing them in selective medium. They are then screened to determine which ones produce antibody of the desired specificity. [After C. Milstein. Monoclonal antibodies. Copyright 1980 by Scientific American, Inc. All rights reserved.]... Figure 4.33. Preparation of Monoclonal Antibodies. Hybridoma cells are formed by fusion of antibody-producing cells and myeloma cells. The hybrid cells are allowed to proliferate by growing them in selective medium. They are then screened to determine which ones produce antibody of the desired specificity. [After C. Milstein. Monoclonal antibodies. Copyright 1980 by Scientific American, Inc. All rights reserved.]...
The next development was the production of monoclonal antibodies (MAbs) in the mid-1970s. This uses hybridoma technology, which involves the fusion of antibody-producing B cells to immortal myeloma cells. Figure 4.4 shows the preparation of MAbs using hybridoma techniques. A more detailed discussion of biopharmaceuticals production is presented in Section 10.5. [Pg.110]

The problem is that if an individual antibody-producing cell is isolated and grown in culture, its descendants have a limited lifespan that severely limits their use for the routine preparation of monoclonal antibodies. In 1975, Milstein and Kohler discovered how monoclonal antibodies of almost any desired antigen specificity can be produced indefinitely and in large quantities. Their method was to fuse a B lymphocyte producing antibody of the desired specificity with a cell derived from a cancerous lymphocyte tumor, called a myeloma cell, which is immortal. The cell fusion is called a hybridoma, which is both immortal and secretes the same specific antibody originally encoded by the B lymphocyte. [Pg.105]

All of the processes and considerations described for polyclonal antibody production are also relevant to monoclonal antibody production. The only variation between the two, with respect to the the immunization process, is species selection. Monoclonal antibodies are most commonly produced in mice because of the commercial availability of mouse myeloma cells that are particularly suited as partner cells in hybridoma preparation (discussed below). [Pg.114]

In fact the preparation of monoclonal antibodies rests on another principle. The reader will realize, as in all other more traditional branches of chemistry, that considerable progress is being made when pure compounds are obtained. As we have seen, after immunization with one antigen (Ag), there is a proliferation of B lymphocytes, each one specialized in the production of one of the anti-Ag antibodies, say X, X, X", etc. If it were possible to cultivate these cells in vitro, these clones could be separated by established methods. Unfortunately, it is not possible to cultivate B lymphocytes in vitro. The problem has been solved by using the properties of a line of myeloma cells. [Pg.131]

An alternative procedure is to isolate the B-cells, fuse them with a tumor line (myeloma), and separate these hybrid cells into individual clones, to produce antibody producing hybrids called hybridomas [10-12]. These separate cell lines can be screened to select a clone that produces an antibody with a single structure, called a monoclonal antibody. For analytical purposes, one advantage of a monoclonal antibody is that the cell line can be preserved so that a consistent source of antibody can be obtained. Again, usually researchers prepare their own monoclonals or obtain cell lines from other investigators who have published reports about the use of their monoclonals. Many researchers deposit their cultures with the American Type Culture Collection, Rockville, MD. [Pg.185]

Passive antibody vaccines have been prepared up to now from human blood serum. Consequently, there has been no need for cultivation methods beyond vaccination and conventional harvest of antibody-containing blood from donors. Due to safety concerns over using human blood, passive vaccines will likely be monoclonal antibodies or cocktails thereof prepared in vitro by the cultivation of hybridoma or myeloma cell lines. This approach is under investigation for anti-HIV-1 antibodies [Emini et al., 1992]. Cultivation of these cell lines involves the same principles of animal cell cultivation as described above, with the exception that hybridomas can be less fastidious in nutritional requirements, and they do not require surface attachment for growth. These features will allow for defined serum-free media and simpler cultivation vessels and procedures. [Pg.210]

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... 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...

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