Recombinant Mouse Anti-HCV E2 Antibody (DAO5) (CAT#: PABC-319)

Figure 1 Mouse Anti-HCV E2 Antibody (DAO5) in SDS-PAGE.

Cross-competition profile of DAO5. (A) Cross-competition and biochemical analysis of the sE2412-715-DAO5 complex. (Left) sE2412-715 and an sE2412-715-DAO5 scFv complex were affinity loaded onto a StrepTactin column, a CBH-4D Fab fragment was added, and the eluted complex was analyzed by SDS-PAGE under nonreducing conditions. (Right) CD81-LEL was incubated overnight at RT with the full-length HCV ectodomain (comprising residues 384 to 715; sE2) and the sE2-DAO5 Fab complex, followed by SEC and analysis of the peak fractions by SDS-PAGE under reducing conditions. DAO5 heavy and light chains form an apparent single band in the reducing gel due to an almost identical molecular mass of ~24 kDa.

Vasiliauskaite I, Owsianka A, England P, et al. Conformational flexibility in the immunoglobulin-like domain of the hepatitis C virus glycoprotein E2[J]. MBio, 2017, 8(3): e00382-17.

Figure 2 Mouse Anti-HCV E2 Antibody (DAO5) in SEC-HPLC.

A preformed sE2412-715-DAO5 Fab complex was incubated in the absence or presence of a Fab fragment targeting a non-overlapping (AP33) or overlapping (e137) region of cE2 and analyzed by SEC. After preincubation with AP33 (green), appearance of a peak at a higher molecular mass indicated ternary complex formation; after preincubation with e137 Fab (blue), the presence of peaks corresponding to the binary complex (at ~13 ml) and an isolated Fab fragment (at ~16 ml) showed that no ternary complex was formed.

Vasiliauskaite I, Owsianka A, England P, et al. Conformational flexibility in the immunoglobulin-like domain of the hepatitis C virus glycoprotein E2[J]. MBio, 2017, 8(3): e00382-17.

Figure 3 Mouse Anti-HCV E2 Antibody (DAO5) in SDS-PAGE.

sE2412-715 or sE2 was immobilized on a StrepTactin column and incubated first with a molar excess of DAO5 scFv and subsequently with e137.

Vasiliauskaite I, Owsianka A, England P, et al. Conformational flexibility in the immunoglobulin-like domain of the hepatitis C virus glycoprotein E2[J]. MBio, 2017, 8(3): e00382-17.

Figure 4 Mouse Anti-HCV E2 Antibody (DAO5) in SPR.

Real-time SPR analysis of Fab binding to immobilized sE2 recorded the binding response (in resonance units [RU]) as a function of time. Fab fragments of e137 and DAO5 were injected over a surface with immobilized HCV sE2.

Vasiliauskaite I, Owsianka A, England P, et al. Conformational flexibility in the immunoglobulin-like domain of the hepatitis C virus glycoprotein E2[J]. MBio, 2017, 8(3): e00382-17.