Rituximab Formulated Bulk (For Further Manufacturing Use) - CD20 monoclonal antibody, recombinant
Status - approved in U.S.; not marketed
Organizations involved:
Genentech, Inc. – Manuf.
Biogen Idec, Inc. – Manuf.
Lonza Biologics plc – Manuf.
Alusuise-Lonza Group – Parent
IDEC Pharmaceuticals Corp. – Former
Columbia University – Tech.; Patent dispute
Amgen – Tech.
Cross ref.: See the entry above for Rituximab (Rituxan), the finished product manufactured from this bulk intermediate. See the entry for Monoclonal Antibodies (#300).
Description: Rituximab Formulated Bulk (For Further Manufacturing Use) is a bulk (not final or formulated) form of recombinant chimeric murine/human monoclonal antibody glycoprotein targeted to the CD20 receptor on B-cells expressed by a Chinese hamster ovary (CHO) cell line, manufactured by Biogen Idec, Inc. exclusively for further manufacture of formulated rituximab (Rituxan) by Genentech, Inc. Rituximab is produced by CHO suspension culture in a culture medium containing the antibiotic gentamicin, with purification including affinity and ion exchange chromatography. The purification processes include specific viral inactivation and removal procedures (see the Manufacturing section).
Rituximab is an IgG1 kappa immunoglobulin containing murine light- and heavy- chain variable region sequences and human constant region sequences. The monoclonal antibody is targeted to the CD20 receptor antigen expressed on the surface of mature B-lymphocytes (B-cells) and B-cell-derived tumor cells. Rituximab is composed of two heavy chains of 451 amino acids and two light chains of 213 amino acids (based on cDNA analysis). The molecular weight is 144,544 Daltons (155.544 kDa), based on the primary sequence of the reduced, non-glycosylated form. Three oligosaccharide structures (G0, G1 and G2 glycoforms) have been identified by capillary zone electrophoresis (CE) and MALDI-TOF, with no evidence for a terminal sialic acid-containing oligosaccharide. See the Rituxan entry above for further information.
Nomenclature: CD20 Mab, rDNA conc. [BIO]; Rituxan [TR]; Rituximab Formulated Bulk (For Further Manufacturing Use) [FDA]; anti-CD20 monoclonal antibody [SY]; CD20 monoclonal antibody, recombinant [SY]; ; immunoglobulin G 1 (human-mouse monoclonal IDEC-C2B8 gamma1-chain anti-human antigen CD 20), disulfide with human-mouse monoclonal IDEC-C2B8 kappa-chain, dimer [CAS]; anti-(human CD20 antigen) immunoglobulin G1 (human-mouse monoclonal IDEC-C2B8 gamma1-chain) disulfide with human-mouse monoclonal IDEC-C2B8 gamma-chain, dimer [CAS]; 174722-31-7 [CAS RN]; anti-CD20 monoclonal antibody [SY]; IDEC-C2B8 [SY]; pan-B antibodies [SY]; CD20 monoclonal antibody, recombinant [SY]
Companies.: Rituximab was discovered and initially developed by IDEC Pharmaceuticals Corp., now Biogen Idec. Rituxan was commercially developed by a collaboration of IDEC, Genentech, F. Hoffmann-La Roche, Ltd. (majority owner of Genentech) and Zenyaku Kogyo Co., Ltd. See the Rituxan entry above for further information about corporate relationships.
As originally approved, Rituximab Formulated Bulk (For Further Manufacturing Use) was manufactured by IDEC Pharmaceuticals Corp. (now Biogen Idec) at facilities in La Jolla, CA, CBER/FDA est. no. 1235, and further processed, formulated and packaged by Genentech, Inc., CBER/FDA lic. no. 1048. Subsequently, Genentech received sBLA approval for large-scale manufacture of bulk rituximab at its South San Francisco facility to supplement bulk rituximab manufactured by IDEC. In early 1998, Genentech received FDA approval for manufacture using a 12,000 L scale bioreactor. IDEC transferred most bulk rituximab manufacturing responsibilities to Genentech in the 3rd quarter of 1999. In mid-2005, press coverage of Genentech’s manufacturing activities and capacity no longer mentions Biogen Idec as a manufacturer of rixuximab bulk. IDEC/Biogen Idec had been manufacturing rituximab using a 3,000 L cell culture bioreactor. Biogen Idec and Genentech both held biologics licenses for rituximab manufacture, and have described their involvement as co-Manufacturers. Biogen Idec was building a 210,00 sq. ft plant including a 10,000 L bioreactor in Mountain View, CA, which was planned to assume all bulk rituximab manufacture upon completion and FDA approval.
Genentech received approval for rituxan manufacture at new facilities in Vacaville, CA, in April 2000; and in March 2004, reported that Rituxan was being manufactured at this site using larger-scale equipment.
in April 2004, Genentech concluded a long-term manufacturing contract with Lonza Biologics plc for manufacture of rituximab bulk at its facilities in Portsmouth, NH, with Lonza replacing Biogen Idec as contract manufacturer. Although Genentech does not report the distribution of manufacturing capacity between itself and contract manufacturers, it has reported that it expects Lonza to manufacture about one-half of rituxan bulk by late 2005 (after expected FDA approval). Genentech reports that it hopes to further expand Rituxan (and also Avastin) manufacture by about 50% using its currently-licensed cell line. FDA sBLA approval for related process changes for Rituxan was expected by late 2006.
Manufacture: Rituximab is produced in batch culture using a plasmid-transformed Chinese hamster ovary (CHO) cell line (apparently TCAE 8 deposited as ATCC 9119) cultured in serum free medium. Purification involves two chromatography steps using Protein A affinity purification and anion exchange chromatography. Immobilized or matrix-bound Protein A has nonspecific affinity for (binds) most antibodies. The anion exchange chromatography steps further remove impurities such as Protein A, host cell DNA, as well as endogenous and adventitious viruses. Limits during the recovery process have been established for product titer, protein step yields, bovine IgG, Protein A, bioburden, and endotoxin to assure consistency. Bioburden, rodent parvovirus PCR, and mycoplasma PCR are tested at the end of the production run. Bulk product may be stored frozen at -20˚C for up to 18 months. Frozen formulated bulk is thawed and sterile filtered prior to aseptic filling by Genentech.
Process changes have been implemented to alleviate European Union and other concerns regarding use of bovine and human raw materials. Autoclaving of media components at 121˚C or higher has been added. Transferrin (bovine-derived) is no longer used in the culture medium, and has been replaced by an increased amount of ferrous sulfate already present in the culture medium. A replacement Master Cell Bank (MCB) was developed because of inadequate supply of initial cell bank vials. The MCBs have passed various biochemical, biological and immunological tests to assess cell line identity and freedom from adventitious agents. Both the initial MCB and Working Cell Bank (WCB) are frozen in a medium containing 95% fetal bovine serum (FBS). The FBS has been tested for bovine viruses (BVD, PI-3, IRB). Bovine source materials comply with European Union regulations concerning potential BSE/TSE contamination. Gentamicin (gentamycin) is now added to the culture medium as a prophylactic measure against mycoplasma contamination.
Certificates of analysis with predefined specifications are used at the following points: the pre-harvested cell culture fluid (show no adventitious viral or mycoplasmal contamination); the formulated bulk (tested for identity, purity, potency, strength and bioburden); and upon receipt of bulk by Genentech. A certificate of analysis prepared by Genentech includes testing for sterility (meets European Pharmacopoeia), identity, endotoxin, and protein content. Following final formulation of the bulk product, analyses for pH, polysorbate 80 concentration, and sodium concentration are performed, and pH and osmolality of the final dosage form are confirmed.
See also discussion of manufacturing in the Companies section above.
FDA class: Biologic BLA
CBER class: Blood And Blood Derivatives
CBER to CDER: Among the products transferred within FDA on June 30, 2003
Approvals: Date = 19971126; first approval, BLA, granted to IDEC Pharmaceuticals Corp.
Indications: [based n approval letter] for manufacture of concentrated bulk rituximab exclusively provided to Genentech, Inc. for further manufacture of the formulated product, Rituximab
Status: Rituximab Formulated Bulk (For Further Manufacturing Use) is exempt from FDA CBER lot release requirements (as is the finished product, Rituxan).
Tech. transfer: See the Rituxan entry above for patents and technology transfer information.
Companies involvement:
Full monograph
116 CD20 Mab, rDNA conc.
Nomenclature:
CD20 Mab, rDNA conc. [BIO]
Rituxan [TR]
Rituximab Formulated Bulk (For Further Manufacturing Use) [FDA]
Anti-(human CD20 antigen) immunoglobulin G1 (human-mouse monoclonal IDEC-C2B8 ?1-chain) disulfide with human-mouse monoclonal IDEC-C2B8 ?-chain, dimer [CAS]
immunoglobulin G 1 (human-mouse monoclonal IDEC-C2B8 gamma1-chain anti-human antigen CD 20), disulfide with human-mouse monoclonal IDEC-C2B8 kappa-chain, dimer [CAS]
174722-31-7 [CAS RN]
anti-CD20 monoclonal antibody [SY]
CD20 monoclonal antibody, recombinant [SY]
IDEC-C2B8 [SY]
MabThera [TR used by Roche outside of U.S. and Japan]
FDA Class: Biologic BLA
Year of approval (FDA) = 1997
Date of 1st FDA approval = 19971126
(in format YYYYMMDD)
Index Terms:
antibodies (see also immune globulins; monoclonal antibodies)
biopharmaceutical products
exempt from CBER lot release requirements
hamster source materials
intermediate/precursor products
monoclonal antibodies
monoclonal antibodies, recombinant
recombinant DNA
rodent cells <!-- rodentcells -->
Chinese hamster ovary (CHO) cells
glutamine synthetase (GS) expression system
mammalian cell culture
rodent cells <!-- rodentcells -->
Protein A affinity chromatography
approval dates uncertain (FDA reports erroneous, conflicting, or simply has lost the original approval dates) (FDAapproved)
approval dates uncertain (FDA reports erroneous, conflicting, or simply has lost the original approval dates) (FDAapproved)
catheter clearance
exempt from CBER lot release requirements
North American coral snake
EU000 Not yet/Never filed with EU
UM999 Not Available/Not Marketed in US
US200 Currently Approved in US
EM999 Not Available/Not Marketed in EU
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