Development of antiricin single domain antibodies toward detection and therapeutic reagents.
Single domain antibodies (sdAb) that bind ricin with high affinity and specificity were selected from a phage display library derived from the mRNA of heavy chain antibodies obtained from lymphocytes of immunized llamas. The sdAb were found to recognize three distinct epitopes on ricin. Representative sdAb were demonstrated to function as both capture and tracer elements in fluid array immunoassays, a limit of detection of 1.6 ng/mL was obtained. One sdAb pair in particular was found to be highly specific for ricin. While polyclonal antibodies cross react strongly with RCA120, the sdAb pair had minimal cross reactivity. In addition, the binders were found to be thermal stable, regaining their ricin binding activity following heating to 85 degrees C for an hour. Cycles of thermally induced unfolding of the sdAb and their subsequent refolding upon cooling was monitored by circular dichroism. As several of the sdAb were observed to bind to ricin's A chain, cell free translation assays were performed to monitor the ability of the sdAbs to inhibit ricin's biological activity. One of the sdAb (C8) was particularly effective and blocked ricin's biological activity with an effectiveness equal to that of a mouse antiricin antibody. These results indicate that antiricin sdAb have great potential for both diagnostic and therapeutic applications.
Anderson
George P
GP
Center for Bio/Molecular Science and Engineering, Naval Research Laboratory, 4555 Overlook Avenue SW, Washington, DC 20375, USA.
Liu
Jinny L
JL
Hale
Martha L
ML
Bernstein
Rachael D
RD
Moore
Martin
M
Swain
Marla D
MD
Goldman
Ellen R
ER
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D002161:Camelids, New World / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D049109:Cell Proliferation; D002619:Chemical Warfare Agents / Q000494:pharmacology*; D002942:Circular Dichroism; D007118:Immunoassay; D007143:Immunoglobulin Heavy Chains / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D008156:Luciferases / Q000378:metabolism; D008214:Lymphocytes / Q000276:immunology; D051379:Mice; D008863:Microspheres; D019151:Peptide Library; D014176:Protein Biosynthesis; D012276:Ricin / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D013945:Thymoma / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D013953:Thymus Neoplasms / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D014407:Tumor Cells, Cultured
D000906:Antibodies; D002619:Chemical Warfare Agents; D007143:Immunoglobulin Heavy Chains; D019151:Peptide Library; D012276:Ricin; D008156:Luciferases