Correction: Quantitative Assessment of Antibody Internalization with Novel Monoclonal Antibodies against Alexa Fluorophores.

There are a number of errors in the headings for Fig 1B, “Quenching by anti-Alexa Fluor mAbs.” “Beads coated with 1C1-A488” and “Beads stained with 1C1-A488” should be “Beads coated with 1C1-A594” and “Beads stained with 1C1-A594”. Please see the corrected Fig 1 here.

Fig 1

Quenching by anti-Alexa Fluor mAbs.

(A) Fluorescence of Alexa Fluor 488 (A488) on microbeads coated with 1C1-A488 or PC-3 cells stained with 1C1-A488 was quenched with a titration of the benchmark, a rabbit anti-A488 polyclonal, or 1 of 3 anti-A488 mAbs. One representative experiment of multiple is shown. (B) Fluorescence of Alexa Fluor 594 (A594) on microbeads coated with 1C1-A594 or PC-3 cells stained with 1C1-A594 was quenched with a titration of 1 of 3 anti-A594 mAbs. One representative experiment of multiple is shown. (A, B) Median fluorescence intensities (MFIs) at each anti-A488 or anti-A594 mAb concentration were normalized against a buffer control. The chimeric IgG1 isotype control was used as a non-quenching mAb control. The IC50 values (microgram/ml) of quenching and the corresponding 95% confidence intervals (95% CI) are listed for both the microbead- and cell-based titrations.

There are errors in the equation in the second to last sentence of the penultimate paragraph of the subsection “Flow cytometry-based internalization assay using cell surface fluorescence quenching.” The correct sentence should be: Internalized fluorescence was calculated from quenched and non-quenched sample data by correcting for incomplete surface quenching: 1 – (N1 – Q1)/(N1 – (N1Q0/N0)) with N1 = Unquenched MFI at each time point (t1); Q1 = Quenched MFI at t1; Q0 = Quenched MFI for the sample kept on ice (t0); N0 = Unquenched MFI at t0. The publisher apologizes for the error.

There are errors in the equation in the first sentence of the last paragraph of the subsection “Flow cytometry-based internalization assay using cell surface fluorescence quenching.” The correct sentence should be: Internalized fluorescence was plotted and a curve fit was obtained in Prism by nonlinear regression with the one-phase association equation: Y = Y0 + (Plateau – Y0) * (1 – exp(-K*x)) with Y0 = 0 when X (time) is zero and Plateau = 100. The publisher apologizes for the error.