Rabbit monoclonal Fab derived from a phage display library.
Rabbit monoclonal antibodies (RmAb) are not routinely obtained by eukaryotic cell fusion techniques. Therefore, we have applied phage display technology to produce a recombinant rabbit Fab molecule directed against the KLH model antigen. The Fab fragments selected from the rabbit phage display library were subcloned in an expression vector to permit the production of a fusion protein comprising a dimer of bacterial alkaline phosphatase (phoA). This fusion protein was directly produced into the periplasmic space of Escherichia coli. We show that a crude extract containing these conjugates can be used in a direct enzyme immunoassay, as exemplified in the case of the KLH antigen.
Foti
M
M
CNR Cellular and Molecular Pharmacology Center, Milano, Italy. mariaf@farma.csfic.mi.cnr.it
Granucci
F
F
Ricciardi-Castagnoli
P
P
Spreafico
A
A
Ackermann
M
M
Suter
M
M
D000469:Alkaline Phosphatase / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D000918:Antibody Specificity; D001435:Bacteriophages / Q000235:genetics; D001483:Base Sequence; D004797:Enzyme-Linked Immunosorbent Assay; D006433:Hemocyanins / Q000276:immunology; D007124:Immunoenzyme Techniques; D007128:Immunoglobulin Fragments / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics; D008969:Molecular Sequence Data; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism
D000911:Antibodies, Monoclonal; D007128:Immunoglobulin Fragments; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D011993:Recombinant Fusion Proteins; D006433:Hemocyanins; D000469:Alkaline Phosphatase; C032808:keyhole-limpet hemocyanin