9 After this chromatography, incubation in 0.1 molar (M) citric acid pH 3.0 for an hour at 4☌ was carried out. Purification consists of several chromatography gel filtration cycles in Sephadex G-25 coarse (Amershan-Pharmacia, Uppsala, Sweden) Protein an affinity. 8 Media and buffer preparation is performed manually with powders in mixing tanks. Subsequently, 10 6 hybridoma cells are inoculated intraperitoneally in each mineral oil primed BALB/c mice. Justification of this limit should include information concerning the yield of monoclonal antibodies and the stability of the hybridoma. The maximum permissible number of serial passages in culture during normal production is defined and restricted. Each production batch is originated from a fresh ampoule of the seed (e.g., working cell bank). Inoculum preparation includes maintaining the Master and Working Cell Banks (MCB/MWCB) and growing the suspended or stationary methods in spinner flasks. Flowchart of the Immunosorbent CB.Hep-1 Production and Controls All data generated during this process has been analyzed using the Microsoft Excel software version 7.0. In every step, several unitary operations, processes, and quality controls are carried out. Conformation of the cell banks and inoculum The production process of MAb CB.Hep-1 includes four main steps:ġ. In this paper we introduce and apply applicable GMP elements and a documentation system that supports process consistency in the production of immunosorbent CB.Hep-1.ĬB.HEP-1 MONOCLONAL ANTIBODY (ANTI-HBSAG) IMMUNOAFFINITY MATRIX 6 Process validation is an important tool for the evaluation of biotherapeutic products, such as MAbs obtained from mice. In both cases, the starting material is capable of harboring adventitious agents - generally viruses that can result in significant harm to humans. The main characteristic of these products is that they are produced by mammalian cells in culture or are taken directly from an animal. 3 The resulting hybrid cells or hybridomas can be maintained in vitro or in vivo to continuously secrete large quantities of MAb with a defined specificity and can be purified and used in a variety of applications. The technique of Köhler and Milstein involves immortalizing antibody-producing plasma cells from an immunized mouse by fusion with myeloma cells (a plasma cell tumor line). These variables demand that any documentation system provide all possible information about the history of the product from the very beginning of the production process and must fulfill all regulatory requirements for these kinds of products. During the production, control, and administration of any pharmaceutical product, special care must be taken because of the many chemical biological substances, live organisms (cells), and various production processes that could be involved. The documentation system is the essential part of any quality system therefore, it should be based on the GMPs and must comply with all relevant aspects for the production and commercialization of the products. Confor-mance to GMPs is basic for state-of-the-art quality systems. 1 These norms establish requirements for the production of drugs for human use on both medium and large-scale manufacture. The GMPs also minimize unforeseen risks that may occur during the screening of the final product. Several issues that may be useful in developing a regulatory strategy for monoclonal antibodies used as reagents are discussed.Ī Good Manufacturing Practice (GMP) system constitutes an associated group of norms and activities destined to guarantee that every product meets and retains the characteristics of design required for its use. The principal recommendation in this paper is the application of the MAb as a biological reagent in the drug substance manufacturing process where the MAb is used to purify the drug substance. Starting in 1991, the department responsible for immunosorbents CB.Hep-1 production has followed Good Manufacturing Practices (GMPs) regulations by establishing a complete documentation system, in which the main objective has been to guarantee stable and solid production processes for the preparation of biopharmaceutical products. Considering the application of this MAb and issues raised by regulatory agencies for the manufacture of biological products, a documentation system and methods to control the MAb production process were established. This MAb is routinely used for the immunopurification of recombinant Hepatitis B surface Antigen (rHBsAg) for vaccine purposes. The mouse monoclonal antibody (MAb) known as CB.Hep-1 is secreted by the hybridoma 48/1/5/4, specific for the "a" determinant of the Hepatitis B surface antigen (HBsAg).
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