Biomaterial Database

Single-Domain Antibodies for Intracellular Toxin Neutralization. 2022

Timothy F Czajka, and Nicholas J Mantis

Department of Biomedical Sciences, University at Albany School of Public Health, Albany, NY, USA.

Ricin is a plant-derived toxin with a history as a biothreat agent. The toxin's enzymatic subunit, ricin toxin A chain (RTA), is a ribosome-inactivating protein that, when delivered into the cytoplasm of mammalian cells, arrests protein synthesis with extraordinary efficiency. Once within the cytoplasm, RTA is shielded from circulating toxin-neutralizing antibodies. Here, we describe methods we developed to neutralize RTA within the cytoplasm of Vero cells using DNA-based delivery of alpaca-derived single-domain antibodies (VHHs) targeting RTA's active site. We describe the design of the VHH expression vectors, assessment of transient expression of VHHs in Vero cells by enzyme-linked immunosorbent assay and western blotting, and cytotoxicity studies. While the protocols here are specific to ricin, they are easily modified for other toxins or even intracellular pathogens such as viruses.

UI	MeSH Term	Description	Entries
D002161	Camelids, New World	Camelidae of the Americas. The extant species are those originating from South America and include alpacas, llamas, guanicos, and vicunas.	Alpacas,Guanacos,Llamas,Lama glama,Lama glama guanicoe,Lama guanicoe,Lama pacos,Vicugna pacos,Vicugna vicugna,Vicunas,Alpaca,Camelid, New World,Guanaco,Llama,New World Camelid,New World Camelids,Vicuna
D002522	Chlorocebus aethiops	A species of CERCOPITHECUS containing three subspecies: C. tantalus, C. pygerythrus, and C. sabeus. They are found in the forests and savannah of Africa. The African green monkey is the natural host of SIMIAN IMMUNODEFICIENCY VIRUS and is used in AIDS research.	African Green Monkey,Cercopithecus aethiops,Cercopithecus griseoviridis,Cercopithecus griseus,Cercopithecus pygerythrus,Cercopithecus sabeus,Cercopithecus tantalus,Chlorocebus cynosuros,Chlorocebus cynosurus,Chlorocebus pygerythrus,Green Monkey,Grivet Monkey,Lasiopyga weidholzi,Malbrouck,Malbrouck Monkey,Monkey, African Green,Monkey, Green,Monkey, Grivet,Monkey, Vervet,Savanah Monkey,Vervet Monkey,Savannah Monkey,African Green Monkey,Chlorocebus cynosuro,Green Monkey, African,Green Monkeys,Grivet Monkeys,Malbrouck Monkeys,Malbroucks,Monkey, Malbrouck,Monkey, Savanah,Monkey, Savannah,Savannah Monkeys,Vervet Monkeys
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D012276	Ricin	A protein phytotoxin from the seeds of Ricinus communis, the castor oil plant. It agglutinates cells, is proteolytic, and causes lethal inflammation and hemorrhage if taken internally.	Castor Bean Lectin,Lectin, Castor Bean,Lectin, Ricinus,Ricin Toxin,RCA 60,RCA60,Ricin A Chain,Ricin B Chain,Ricin D,Ricin I,Ricinus Toxin,A Chain, Ricin,B Chain, Ricin,Ricinus Lectin,Toxin, Ricin,Toxin, Ricinus
D014709	Vero Cells	A CELL LINE derived from the kidney of the African green (vervet) monkey, (CHLOROCEBUS AETHIOPS) used primarily in virus replication studies and plaque assays.	Cell, Vero,Cells, Vero,Vero Cell
D057134	Antibodies, Neutralizing	Antibodies that reduce or abolish some biological activity of a soluble antigen or infectious agent, usually a virus.	Neutralizing Antibodies,Antibody, Neutralizing,Neutralizing Antibody
D061905	Single-Domain Antibodies	An immunoglobulin fragment composed of one variable domain from an IMMUNOGLOBULIN HEAVY CHAIN or IMMUNOGLOBULIN LIGHT CHAIN.	Single-Domain Antibody,VHH Antibodies,VHH Antibody,Immunoglobulin VH Domain Fragments,Immunoglobulin VL Domain Fragments,Nanobodies,VH Domain Fragments,VHH Fragments,VHH Immunoglobulin Fragments,VL Domain Fragments,VNAR Fragments,VNAR Immunoglobulin Fragments,Antibody, Single-Domain,Antibody, VHH,Single Domain Antibody