In this report, we describe the high-yield secretory expression of the recombinant human anti-HBsAg Fab fragment from Pichia pastoris that was achieved by co-integration of the genes encoding the heavy and light chains (both under the control of alcohol oxidase promoter) into the genome of the yeast cells. The fed-batch fermentations were carried out in a 5 L scale. Both chains of the Fab were ...

The Fab'-fragment of a mouse IgA-myeloma (protein-315) was split by pepsin to yield a smaller fragment that retained the anti-2,4-dinitrophenyl activity of the intact protein. This fragment, which we call Fv, has a molecular weight of about 30,000 (half that of Fab'), and is composed of two polypeptide chains (molecular weight 14,000) held together by noncovalent bonds. The N-terminal sequence ...

A copper l immunoglobulin complex isolated from a myeloma patient with hypercupremia has been investigated in an attempt to localize and characterize the copper-binding site. Cleavage of the purified IgGl type immunoglobulin with papain yielded peptides which were identified as Fab and F, fragments by immunodiffusion. The fragments were separated by DEAE-cellulose chromatography and the Fab fra...

Chemical conjugation of small recombinant proteins with polyethylene glycol (PEG) is an established strategy to extend their typically short circulation times to a therapeutically useful range. We have investigated the production of a genetic fusion with a glycine-rich homo-amino-acid polymer (HAP) as an alternative way to attach a solvated random chain with large hydrodynamic volume. The anti-...

Venomous snakebite is a life-threatening injury in many tropical and subtropical areas including Iran. The gold standard treatment option for human envenomation is the use of antivenoms. Despite the unique effects of horse-derived antivenoms on the treatment of snakebite, they are not fully perfect and need improvements. In this study, human recombinant Fab fragment antivenom was produced in Ro...

A mouse monoclonal antibody (mAb) prepared by hybridoma technology could recognize the Cucumber mosaic virus (CMV) coat protein. The Fab fragments (antigen-binding site, one complete light chain, and part of one heavy chain) genes encoding the light chain and the Fd (a monovalent antigen-binding fragment consisting of a complete light chain paired with the variable region and the first constant...

The cDNAs for the two variable domains of the antibody 9E10 were cloned from the hybridoma cell line. A chimeric 9E10 Fab fragment was produced in E. coli under control of the tightly controlled tetracycline promoter. The functional Fab fragment was isolated in a single step via a His6-tag, which also served for its recognition by a nickel chelate-alkaline phosphatase conjugate. Thus, the recom...

CD30, the so-called Reed-Sternberg antigen, constitutes a promising cell-specific target for the treatment of Hodgkin's lymphoma. Starting from the previously characterized cognate HRS3 mouse monoclonal antibody, the bacterially produced functional Fab fragment was humanized by grafting the CDRs from the mouse antibody framework on to human immunoglobulin consensus sequences. This procedure led...

In vitro selection by display methods has been an effective tool for engineering recombinant antibodies. mRNA display based on a cell-free translation system has the advantages of larger library sizes and quicker selection procedures compared with cell-based display methods such as phage display. However, mRNA display has been limited to select single-chain polypeptides such as scFvs due to its...

In this report, we describe the high-yield secretory expression ( approximately 40 mg x l(-1)) of pure, atrazine-specific Fab fragments (K411B) from Pichia pastoris that was achieved by co-integration of the genes encoding the heavy and light chains (both under the control of the alcohol oxidase promoter) into the genome of the yeast cells. Antibody-expressing clones were selected by SDS-PAGE a...