Ricin is a toxin that could potentially be used as a bioweapon. We identified anti-ricin A chain antibodies by sequencing the antibody repertoire from immunized mice and by selecting high affinity antibodies using yeast surface display. These methods led to the isolation of multiple antibodies with high (sub-nanomolar) affinity. Interestingly, the antibodies identified by the 2 independent approaches are from the same clonal lineages, indicating for the first time that yeast surface display can identify native antibodies. The new antibodies represent well-characterized reagents for biodefense diagnostics and therapeutics development.
Submitter: Marie Ary
Submission Date: Oct. 29, 2018, 11:12 a.m.
|Number of data points||49|
|Proteins||Anti-ricin chimeric Ab ; Anti-ricin scFv ; Anti-ricin scAb|
|Assays/Quantities/Protocols||Experimental Assay: ELISA EC50 ; Experimental Assay: k_off ; Experimental Assay: k_on ; Experimental Assay: Kd - SPR ; Experimental Assay: Kd - yeast display|
|Libraries||Kd, kon, koff by SPR of scAbs from Abs isolated from bone marrow or spleen (Table 2) ; Kd by yeast display of scFvs derived from Abs isolated from bone marrow and spleen (Table 2) ; Binding kinetics of RAM1.5 Ab from draining lymph node by ELISA and SPR ; Binding kinetics RAM1.4 Ab from draining lymph node by ELISA and SPR ; SP1 scAb Kd, k_on, k_off by SPR (Table 2) ; Kd by yeast display of SP1 scFv derived from Abs isolated from bone marrow and spleen (Table 2)|