To develop very small antibody-derived recognition units for experimental, medical, and drug design purposes, a heavy chain variable region (VH) single-domain phage-display library was designed and constructed. The scaffold that was used for library construction was a native sequence of a monoclonal antibody with a unique VH/VL interface. There was no need to modify any residues in the VL interface to avoid non-specific binding of VH domain. The library repertoire, consisting of 4x10(8)independent clones, was generated by the randomization of nine amino acid residues in complementary determining region 3. The library was screened by binding to protein antigens, and individual clones were isolated. The VH genes encoding for specific binding clones were rescued and large amounts of soluble and stable single-domain VH protein were made from insoluble inclusion bodies by in vitro refolding and purification. Biochemical and biophysical characterization of the VH protein revealed a highly specific, correctly folded, and stable monomeric molecule. Binding studies demonstrated an affinity of 20 nM. The properties of these molecules make them attractive for clinical, industrial, and research applications, as well as a step toward improvement in the design of small molecules that are based on the hypervariable loops of antibodies.

Download full-text PDF

Source
http://dx.doi.org/10.1006/jmbi.1999.2923DOI Listing

Publication Analysis

Top Keywords

heavy chain
8
chain variable
8
variable region
8
library
5
antibody single-domain
4
single-domain phage
4
phage display
4
display library
4
library native
4
native heavy
4

Similar Publications

The global prevalence of heart failure is still growing, which imposes a heavy economic burden. The role of microRNA-146b (miR-146b) in HF remain largely unknown. This study aims to explore the role and mechanism of miR-146b in HF.

View Article and Find Full Text PDF

Chronic lymphocytic leukemia (CLL) is a less common hematological malignancy in Indian people. It accounts for less than 5% of all leukemias. Information on genomic alteration in CLL is limited immunoglobulin heavy-chain variable region (IGHV) mutational status is considered the most reliable prognostic marker.

View Article and Find Full Text PDF

Nanobodies or variable antigen-binding domains (VH) derived from heavy chain-only antibodies (HcAb) occurring in the Camelidae family offer certain superior physicochemical characteristics like enhanced stability, solubility, and low immunogenicity compared to conventional antibodies. Their efficient antigen-binding capabilities make them a preferred choice for next-generation small biologics. In the present work, we design an anti-SARS-CoV-2 bi-paratopic nanobody drug conjugate by screening a nanobody database.

View Article and Find Full Text PDF

The increasing level of cadmium (Cd) contamination in soil due to anthropogenic actions is a significant problem. This problem not only harms the natural environment, but it also causes major harm to human health via the food chain. The use of chelating agent is a useful strategy to avoid heavy metal uptake and accumulation in plants.

View Article and Find Full Text PDF

PbOI: A Lead Oxyhalide IR Optical Crystal with an Unprecedented [OPb] Chain Featuring a Wide Transmittance Range and Large Birefringence.

Inorg Chem

January 2025

Research Center for Crystal Materials, State Key Laboratory of Functional Materials and Devices for Special Environmental Conditions, Xinjiang Key Laboratory of Functional Crystal Materials, and Xinjiang Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, 40-1 South Beijing Road, Urumqi 830011, P. R. China.

Among the infrared (IR) optical material systems, the heavy-metal oxyhalide system has become an emerging system in recent years. Introducing heavy-metal cations and halogen anions with large atomic numbers is conducive to widening the IR transparency window and improving the birefringence value. Our experiments focus on the PbO-PbI system and find a new lead oxyhalide, PbOI.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!