Skip to main content
. Author manuscript; available in PMC: 2015 Dec 1.
Published in final edited form as: Arthritis Rheumatol. 2014 Dec;66(12):3445–3456. doi: 10.1002/art.38872

Figure 2. Labial salivary gland-derived human recombinant antibodies produced by an SS/SLE Patient have specificity to Ro, La and SmRNP.

Figure 2

Recombinant human monoclonal antibody screening for affinity to Sm (■), SmRNP (▲), Ro (◆) and La (▼) by ELISA (Positive cutoff ≥ 0.5 OD405). High affinity binding of A2-G03 anti-Ro by ELISA (A, left panel; Kd=7×10−10M) and A3-E03 to SmRNP (A, right panel; Kd=9.0×10−11M). INNO-LIA™ analysis of A2-G03 anti-Ro antibody to both 52 kD and 60kD Ro antigens (B, center strip), and SmB by A2-G05 antibody (B, right strip). Anti-nuclear antibody staining for A2-G03 (C, top panel) and A3-E03 anti-SmRNP antibody (C, bottom panel). 35S protein immunoprecipitation (35S IP) and RNA co-immunoprecipitation (RNA IP) of HeLa cell lysate (D) for A2-G03 to 60 kD Ro (35S IP, 3rd from left) and proteins from the SmRNP complex by A3-E03 (35S IP, 4th from left, arrows A and B/B′). RNA IP analysis showed co-precipitation of hYRNA3, hYRNA4 and hYRNA5 by A2-G03 (RNA IP, 2nd from left) and of multiple U-Small nuclear RNP complexes, particularly U1RNP by A3-E03 (RNA IP, 3rd from left). A U3-Ro standard (RNA IP, left) serum and total nucleic acids (TNA) (RNA IP, right) were included as controls.