Systematic generation of antigen specific human monoclonal antibodies with therapeutical activities using active immunization

Affinity Constants of Naturally Acquired and Vaccine-Induced Anti-Pseudomonas aeruginosa Antibodies in Healthy Adults and Cystic Fibrosis Patients

“…The specificity of this assay system was verified with monoclonal antibodies specific for P. aeruginosa LPS [17,18]. Only antibodies specific for IATS 6 LPS or those that recognize antigenic determinants common to most serotypes of P. aeruginosa LPS gave a signal in this assay system.…”

“…Serum antibodies specific for LPS were quantitated by ELISA as described elsewhere [5,17,18]. For coating, purified LPS (5 mg/rnl) dissolved in 36 mM triethylamine was used.…”

Scale-up and optimization of culture conditions of a human heterohybridoma producing serotype-specific antibodies to Pseudomonas aeruginosa

“…The heterohybridoma (human x human xmouse) used in this study, designated 4-8KH15, secretes serotype specific human monoclonal IgM specific for P. aeruginosa LPS of Habs serotype 4. Isolation of antigen-specific lymphocytes, Epstein-Barr Virus (EBV) transformation and fusion with a heteromyeloma have been described previously (Lang et al 1989a(Lang et al , 1989b(Lang et al , 1990. Cells were cultured in HEPES-buffered Iscove's modified Dulbecco's medium (IMDM; Sigma, St. Louis, Mo., USA) supplemented with 2% foetal bovine serum, 5 x 10-SM 2-mercaptoethanol and 35 mM Nat-ICO3.…”

“…Out of a panel of heterohybridomas secreting serotype specific human IgM MAb against lipopolysaccharides (LPS) of the Gram-negative bacterial strain Pseudomonas aeruginosa and against capsular polysaccharides of Klebsiella pneumoniae (Lang et al 1989a(Lang et al , 1989b(Lang et al , 1990(Lang et al , 1991a(Lang et al , 1991b, one hybridoma cell line (4-8KH15) was selected to investigate the influence of pH and growth rate on antibody production under controlled culture conditions. The cell line 4-8KH15 is a representative sample of the whole panel of hybridomas.…”

Mining human antibody repertoires

“…Today rodent mAbs against a plethora of antigens are widely involving EBV-immortalization of peripheral B cells followed by fusion with myeloma cells and cloning of hybridomas were established. [45][46][47][48] This method has yielded several human mAbs with therapeutic potential, including neutralizing antibodies against the hemagglutinin protein derived from the 1918 H1N1 pandemic influenza virus. 49 As an alternative to the formation of hybridomas, EBVtransformed cells have been infected with a retrovirus encoding the ras oncogene, yielding cells with similar cloning efficiencies and levels of antibody secretion.…”