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Abstract

The protein-free medium TurboDoma HP.1 (THP.1) was used to produce the CB.Hep-1 monoclonal antibody (mAb) in a CP-1000 hollow fiber bioreactor (HFB). This mAb is used for the immunopurification of recombinant hepatitis B surface antigen (rHBsAg), which is included in a vaccine preparation against the Hepatitis B Virus. By using the experimental conditions tested in this work we were able to generate more than 433 mg of IgG in 43 days. The maximum antibody concentration obtained was about 2.4 mg ml(-1)and the IgG production per day was approximately 11 mg of monoclonal antibody, which constitutes a good concentration value in comparison to the results obtained in ascitic fluid, where concentration for this hybridoma was around 3 mg ml(-1). We used different analytical methods to control the quality of mAbs, obtained from the in vitro system. They included affinity constant determination, analysis of N-glycan structures, immunoaffinity chromatography and antigen binding properties. The results obtained suggest that no significant changes occurred in the mean characteristics of the mAb harvested from the bioreactor during the 43 days of cultivation.

Expression and characterization of an anti‐(hepatitis B surface antigen) glycosylated mouse antibody in transgenic tobacco (Nicotiana tabacum) plants and its use in the immunopurification of its target antigen

“…The N‐glycan composition of the plantibody was determined and compared with the N‐glycan structures present in the mouse mAb [50]. The mouse mAb is characterized by a simple and homogeneous glycosylation pattern in which the asialo agalacto‐fucosylated biantennary structure is the most abundant complex N‐glycan, followed by asialo‐monogalacto‐fucosylated biantenna and a very small proportion of the asialo‐digalacto‐fucosylated biantenna.…”

Lyman Alpha Galaxies: Primitive, Dusty, or Evolved?

“…The CTIO data were reduced with IRAF 1 (Tody 1986(Tody , 1993, using the MSCRED (Valdes & Tody 1998;Valdes 1998) reduction package, following the method set forth in Rhoads et al (2000Rhoads et al ( , 2004. First, we performed the standard image reduction steps of overscan subtraction and bias subtraction, followed by flat-fielding using dome flats obtained during our run.…”

Influence of culture conditions on the N‐glycosylation of a monoclonal antibody specific for recombinant hepatitis B surface antigen

“…Variations in production levels were observed (Table 1). However, bioactivity in terms of the affinity constant (3.6×10 8 M −1 ) and immunoaffinity efficiency of immunoabsorbent gel for antigen purification [1025 μg·of rHBsAg·(ml of gel) −1 ] remained almost unchanged, as reported by Valdés et al [13].…”

“…PBS (pH 8.0) and 0.1 M citric acid (pH 4.0) were used for adsorption and elution respectively with a flow rate of 100 cm·h −1 . The affinity constant and immunoaffinity efficiency were determined by the method described previously [13].…”