抗体抗原实验

This protocol describes a method for creation of a highly diverse and functional synthetic phage-displayed repertoire of fully human domain antibodies (dAbs). The repertoire is based on two human frameworks (one VH and one Vκ) that express well in bacteria and are frequently used in human anti ...

Immunoglobulin new antigen receptors (IgNARs) from sharks are a distinct class of immune receptors, consisting of homodimers with no associated light chains. Antigen binding is encapsulated within single VNAR immunoglobulin domains of 13–14 kDa in size. This small size and single dom ...

The immune response of infected or immunized dromedaries contains a diverse repertoire of conventional and heavy chain-only antibodies, both functional in antigen binding. By definition, a heavy chain antibody is devoid of a light chain and in the case of the heavy chain antibodies in cameli ...

The development of therapeutic monoclonal antibodies over the last 35 years has led to the emergence of a new class of useful therapeutic molecules. These “first generation” antibodies have been obtained thanks to the conjugated and huge efforts of both academic and biotech researchers. A ...

In this chapter we describe in detail the current protocols that are used to express single-domain antibodies in bacteria. Bacteria are among the most common expression systems for expressing recombinant proteins. We present different approaches for carrying out periplasmic and cy ...

The production of VHHs in microorganisms is relatively straightforward, however the amount of VHH produced per volume unit can vary substantially from hardly detectable to hundreds of milligrams per liter. Expression in Escherichia coli is more commonly used at initial research phas ...

Camelid single domain antibodies fused to noncamelid Fc regions, also called chimeric heavy chain antibodies (cHCAb), offer great potential as therapeutic and diagnostic candidates due to their relatively small size (80 kDa) and intact Fc. In this chapter, we describe two approaches, li ...

Transgenic plants for the production of high-value recombinant complex and/or glycosylated proteins are a promising alternative for conventional systems, such as mammalian cells and bacteria. Many groups use plants as production platform for antibodies and antibody fragment ...

Antibodies are invaluable macromolecules effectively utilized as detection reagents and therapeutics. Traditionally, researchers have relied upon the entire immunoglobulin molecule, however advances in protein engineering have ushered the use of antibody fragments ...

VHHs usually display high physical and proteolytic stability, but in some cases stability needs to be increased further for their intended applications. The high thermal stability is due to the stable 3D structure of VHHs, which consists of a sandwich of nine beta-strands with a high number of int ...

Solubility and stability are amongst the factors contributing to the therapeutic efficacy of biologics. Human antibody heavy chain variable domains, VHs, are one class of biologics; improving VH biophysical properties is the focus of significant protein engineering efforts. Here, ...

The successful medical application of single domain antibodies largely depends on their functionality. This feature is partly determined by the intrinsic stability of the single domain. Therefore a lot of research has gone into the elucidation of rules to uniformly increase stability ...

Human antibody variable heavy (VH) domains tend to display poor biophysical properties when expressed in isolation. Consequently, the domains are often characterized by low expression levels, high levels of aggregation, and increased “stickiness.” Here, we describe methods that a ...

Camelidae single-domain antibodies (VHHs) represent a unique class of emerging therapeutics. Similar to other recombinant antibody fragments (e.g., Fabs, scFvs), VHHs are amenable to library screening and selection, but benefit from superior intrinsic biophysical properties ...

The understanding of cellular processes and their physiopathological alterations requires comprehensive data on the abundance, distribution, modification and interaction of cellular components. On the one hand, artificially introduced fluorescent fusion proteins p ...

Molecular imaging is a noninvasive method to measure specific biological processes in animal models and patients using imaging. In recent years there has been a tremendous evolution in hardware and software for imaging purposes. This progress has created an urgent need for new labeled tar ...

The deposition of peptides and proteins as amyloid fibrils is a common feature of nearly 50 medical �disorders affecting the brain or a variety of other organs and tissues. These disorders, which include Alzheimer’s disease, Parkinson’s disease, the prion diseases, and type II diabetes, have an ...

High avidity, pentameric, single-domain antibodies, oligomerized through the B subunit of verotoxin, are excellent immunohistochemical reagents. The resulting molecules are termed pentabodies. Here, we describe the immunostaining of tissue sections with ES1, a pentabody r ...

One of the advantages of using recombinant instead of conventional antibodies is that these can be easily manipulated by means of standard molecular biology techniques. Therefore this opportunity can be exploited to prepare fusion constructs composed of VHHs and suitable tags. Accor ...

Chromatin immunoprecipitation (ChIP), followed by microarray hybridization (ChIP-chip) or high-throughput sequencing (ChIP-seq), is becoming a widely used powerful method for the analysis of the in vivo DNA–protein interactions at genomic scale. The success of ChIP largely dep ...