Next Article in Journal
Refolding Technologies for Antibody Fragments
Next Article in Special Issue
Reduced Culture Temperature Differentially Affects Expression and Biophysical Properties of Monoclonal Antibody Variants
Previous Article in Journal / Special Issue
Engineered Bovine Antibodies in the Development of Novel Therapeutics, Immunomodulators and Vaccines
Antibodies 2014, 3(2), 215-231; doi:10.3390/antib3020215

Kinetic Characterization of a Panel of High-Affinity Monoclonal Antibodies Targeting Ricin and Recombinant Re-Formatting for Biosensor Applications

1,3, 2,4, 1,5, 2, 2, 2, 1, 1, 1, 1 and 1,*
1 CSIRO Materials Science and Engineering, Parkville, Victoria 3052, Australia 2 Defense Science and Technology Organization, Fishermans Bend, Victoria 3207, Australia 3 The School of Medicine, Deakin University, Waurn Ponds, Victoria 3216, Australia 4 Bio21 Institute, The University of Melbourne, Parkville, Victoria 3052, Australia 5 The Department of Immunology, Monash University, Clayton, Victoria 3800, Australia
* Author to whom correspondence should be addressed.
Received: 19 February 2014 / Revised: 10 April 2014 / Accepted: 28 April 2014 / Published: 9 May 2014
(This article belongs to the Special Issue Antibody Engineering)
View Full-Text   |   Download PDF [1099 KB, 26 May 2014; original version 9 May 2014]   |  


Ricin is a potent glycoprotein toxin that is structurally composed of two subunits joined via a disulfide bond: a ~30 kDa subunit A (RTA) and a ~32 kDa subunit B (RTB). There are fears of ricin being used as a weapon for warfare and terrorism and, as such, there is an increasing need for the development of immunodiagnostic reagents targeted towards this toxin. This article describes the production and characterization of a panel of six ricin-specific monoclonal IgG antibodies (mAbs), previously selected based upon their ability to inhibit ricin-mediated killing of cultured cells. Subsequent epitope binding analysis using the surface plasmon resonance (SPR) array biosensor (ProteOn XPR36) indicated three distinct, non-competitive binding epitopes (“bins”). The association (ka) and dissociation (kd) rate constants and binding affinities (KD) of each of the mAbs to ricin were also determined by SPR using Biacore T100 instrument. Affinities (KD) ranged from 0.1 nM to 9 nM. We present the coding sequences of the variable domains of the six mAbs, the expression, kinetic and cytotoxicity assays for two recombinant Fab (rFab) fragments and demonstrate a rFab affinity improvement by chain-shuffling. Together, these antibodies and constituent rFabs represent a panel of reagents for high-affinity recognition of ricin with potential national security biosensor applications.
Keywords: monoclonal antibody; ricin; Fab fragment; SPR; epitope binning monoclonal antibody; ricin; Fab fragment; SPR; epitope binning
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

Share & Cite This Article

Further Mendeley | CiteULike
Export to BibTeX |
EndNote |
MDPI and ACS Style

Cummins, M.; Dogovski, C.; Robert, R.; Alderton, M.; Chong, D.; Proll, D.; Pontes-Braz, L.; Raicevic, A.; Hattarki, M.; Nuttall, S.; Dolezal, O. Kinetic Characterization of a Panel of High-Affinity Monoclonal Antibodies Targeting Ricin and Recombinant Re-Formatting for Biosensor Applications. Antibodies 2014, 3, 215-231.

View more citation formats

Related Articles

Article Metrics


[Return to top]
Antibodies EISSN 2073-4468 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert