Identification and development of new monoclonal antibodies and other biological molecules such ad Fab fragments, diabodies and peptides is a key focus of many biotechnology and pharmaceutical companies and academic research department in several therapeutic areas, includi ...

The generation of recombinant antibodies by phage display in high-throughput demands fast downstream technologies and streamlined processes for the identification and initial characterisation of individual binders. Next to standard immunological methods such as enzym ...

In recent years, antibody microarrays have developed into an important tool for proteomics. As a multiplexing technique, they facilitate the highly parallel detection of hundreds of different analytes from very small sample volumes of only a few microlitres. This is combined with high se ...

Hybrid sub-micrometer particles are of great interest and have been extensively applied to tailor the catalytic, electrical, optical, magnetic, mechanical or thermal properties of the material. Especially their application in the field of diagnostics and drug delivery systems is a r ...

In this chapter, we describe the generation of immunoliposomes by means of two different methods applying different scFv formats. The cloning strategies of these scFv variants possessing an additional cysteine residue for site-directted coupling are also presented in this chapter. ...

Single chain Fv (scFv) antibody fragments fused to the human immunoglobulin G (IgG) Fc moiety obtain IgG-like properties, but nevertheless they are still encoded by a single gene. As transient production of scFv-Fc proteins in mammalians can easily achieve milligram amounts, this antibo ...

Plants are attractive systems for the expression of full-length antibodies and also for antibody derivatives such as scFvs, diabodies, camelid VHH and minibodies. For ‘Molecular Pharming’ ER retention of antibody fragments has been turned out to be the method of choice for production in le ...

The pubmed search term “pastoris AND (express OR produced OR expression OR production)” yielded 877 hits in December 2008, dated from 1987 to 2009. At the same time, the search term “pastoris AND (bioreactor OR fed-batch OR continuous OR fermentations OR large-scale OR fermentation OR pilot)” re ...

Today, recombinant protein expression in E. coli is usually considered a routine technology. Nonetheless, problems with soluble expression, rooted in protein aggregation competing with folding, are still frequently encountered. Many antibody fragments from natural sources ...

Antibodies are an important and growing family of approved therapeutic drugs. Antibody fragments of various types as well as more traditional monoclonal antibodies can be produced using E. coli as the production host. This chapter provides protocols for producing antibody fragments ...

The Strep-tag is a nine-residue minimal peptide sequence with intrinsic affinity towards streptavidin. This peptide – or its improved version Strep-tag II – can be fused to recombinant antibody fragments and other proteins in various fashions. The fact that the interaction between the St ...

Today, monoclonal antibodies (mAbs) form the largest category of biopharmaceuticals in clinical trials and their number is expanding rapidly (DataMonitor 2007). The antibodies or functional antibody fragments are being produced in artificial production systems like mammal ...

For many applications antibody fragments like scFv or Fab are an alternative to full size IgGs. These fragments can be produced in bacterial host systems, which are less time and cost intensive than mammalian cell culture, the production host for more complex IgGs. The Gram-negative bacterium ...

Immobilized metal affinity chromatography (IMAC) has become a widely used method for the affinity purification of recombinant proteins and, in particular, of antibody fragments produced in Escherichia coli. IDA-Sepharose and NTA-agarose charged with different divalent cati ...

The generation of antibody fusion proteins for target-directed delivery of cytokines constitutes a promising immunotherapeutic approach. The fusion of recombinant antibodies to members of the TNF-family that differ clearly in their soluble and membrane-bound forms for their r ...

Recombinant antibodies have become a standard component of research, diagnostics, and therapy. In the development of recombinant antibodies – irrespective of the final format – a monovalent construct is virtually always the first protein to be tested. This is due to the fact that essential ...

Intermediate-sized antibody fragments, such as scFv-CH3 (minibodies) and scFv-Fc, retain high avidity for their antigen due to their nature of bivalency. In addition, the incorporation of the Fc region enables the generation of antibody fragments with a variety of biological functions ...

Single-chain Fv fragments exhibiting monovalent binding can be converted into bivalent molecules by reducing the linker connecting the two variable domains to approximately five residues resulting in homodimerization of two polypeptide chains. This protocol describes the c ...

The scFv-chain Fv (scFv) corresponds to the smallest functional binding unit of a classical antibody and constitutes a favourite format for the comparative first evaluation of the recombinant potential of an antibody. In this protocol, a two-step cloning strategy is described followi ...

The non-immunogenic antigen binder has emerged as an important component in the research of cell recognition and targeting with the high degree of specificity inherited from prototypical antibody (Hoogenboom 2005; Holliger and Hudson 2005; Hu et al. 1996; Olafsen et al. 2004; Shahied et al. 20 ...