In vivo research

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In vitro methods using semipermeable-membrane-based systems produce mAb in concentrations often as high as those found in ascitic fluid and are free of mouse ascitic fluid contaminants. Some hybridomas do not grow well in culture or are lost in culture.

In vitro methods generally require the use of FBS, which limits some antibody uses. The use of in vitro methods for mAb production generally requires the use of FBS, which is a concern from the animal-welfare perspective.

The loss of proper glycosylation of the antibody in contrast with in vivo production might make the antibody product unsuitable for in vivo experiments because of increased immunogenicity, reduced binding affinity, changes in biologic functions, or accelerated clearance in vivo.

In general, batch-culture supernatants contain less mAb typically 0. Note that semipermeable-membrane-based systems have been developed that can produce concentrations of mAb comparable with concentrations observed in mouse ascites fluid.

In batch tissue-culture methods, mAb concentration tends to be low in the supernatant; this necessitates concentrating steps that can change antibody affinity, denature the antibody, and add time and expense.

Adequate concentrations of mAb might be obtained in semi permeable-membrane-based systems. Most batches of mAb produced by membrane-based in vitro methods are contaminated with dead hybridoma cells and dead hybridoma-cell products, thus requiring early and expensive purification before study.

In vitro culture methods are generally more expensive than the ascites method for small-scale or medium-scale production of mAb Hendriksen and de Leeuw ; Jackson and others ; Peterson Peavey ; Marx ; Lipman The number of mAb produced by in vitro methods is limited by the amount of equipment that it is practical to have available.

The National Academies Press. The high concentration of the desired mAb in mouse ascites fluid avoids the effects of contaminants in in vitro batch-culture fluid when comparable quantities of mAb are used.

The mouse ascites method avoids the need to teach the antibody producer tissue-culture methods. Disadvantages of Mouse Ascites Method The mouse ascites method involves the continued use of mice requiring daily observation. MAb produced by in vivo methods can contain various mouse proteins and other contaminants that might require purification.

The mouse ascites method can be expensive if immunodeficient mice in a barrier facility must be used. In vivo methods can cause significant pain or distress in mice.

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