Journal: ACS Omega

Article Title: Integration of Magnetic Bead-Based Cell Selection into Complex Isolations

doi: 10.1021/acsomega.7b01427

Figure Lengend Snippet: Characterization of bead–antibody binding. (A) Magnetic bead fluorescence intensity curves generated by varying densities of a fluorescently labeled anti-EpCAM antibody on beads. A 50% maximal binding capacity for each bead type was then identified ( Table S2 ). Dots represent the average of three technical replicates consisting of 100 beads each (300 beads total); error bars represent the standard deviation of the analyzed population. (B) Impact of the bead’s antibody density on target capture. The 50% maximal antibody binding density for M-280s and FlowComps is denoted by their respective vertical dashed line. Points represent the average of three technical replicates; error bars represent the standard deviation; * denotes p < 0.05, which in this case applies to both M-280 and FlowComp.

Article Snippet: For experiments involving release, a similar experimental design was followed, except that once washed, the beads and bound cells were dropped into a release well containing either FlowComp Release Buffer (FlowComp Flexi Kit 11061D, Thermo Fisher) or CELLection release buffer (CELLection Biotin Binder Kit 11533D, Thermo Fisher) prepared according to the manufacturer’s instructions (release volume of 110 μL).

Techniques: Binding Assay, Fluorescence, Generated, Labeling, Standard Deviation

Journal: ACS Omega

Article Title: Integration of Magnetic Bead-Based Cell Selection into Complex Isolations

doi: 10.1021/acsomega.7b01427

Figure Lengend Snippet: Characterization of cell capture. (A) Capture of EpCAM-expressing cell lines (Du145 = low, 22Rv1 = medium, LNCaP = high) by each bead type. Beads are abbreviated as follows: SM = Sera-Mag, FC = FlowComp, CELL = CELLection. (B) Nonspecific capture of PBMCs by each bead type across varying PBMC inputs. (C) Direct vs indirect capture of Du145s from a PBMC background. (D) Resultant purity of the captured target cells from direct and indirect capture of Du145s. In all plots, the bars represent the technical replicate average ( n = 3) with error bars representing the standard deviation.

Article Snippet: For experiments involving release, a similar experimental design was followed, except that once washed, the beads and bound cells were dropped into a release well containing either FlowComp Release Buffer (FlowComp Flexi Kit 11061D, Thermo Fisher) or CELLection release buffer (CELLection Biotin Binder Kit 11533D, Thermo Fisher) prepared according to the manufacturer’s instructions (release volume of 110 μL).

Techniques: Expressing, Standard Deviation

Journal: ACS Omega

Article Title: Integration of Magnetic Bead-Based Cell Selection into Complex Isolations

doi: 10.1021/acsomega.7b01427

Figure Lengend Snippet: Characterization of the release from FlowComp and CELLection beads. (A,B) Release of a fluorescently labeled anti-EpCAM antibody from (A) FlowComp and (B) CELLection beads. Beads were labeled with low, medium, and high levels of antibody and released for the specified time intervals. (A,B) Dots represent the average of three technical replicates with each technical replicate representing the average of 100 beads (total of 300 beads); error bars represent the standard deviation of the technical replicates. (C) Release of 22Rv1s from FlowComp and CELLection beads across time. (D) Capture efficiency of both FlowComp and CELLection beads when used to capture Du145, 22Rv1, and LNCaP cells. (E) Release efficiency of the three cell lines following bead-based capture. (F) Effective capture following the release of the cells. Gray bars represent the population of cells lost during the release process because of the inefficient release. In each plot, bars represent an average of n = 3; error bars represent the standard deviation.

Article Snippet: For experiments involving release, a similar experimental design was followed, except that once washed, the beads and bound cells were dropped into a release well containing either FlowComp Release Buffer (FlowComp Flexi Kit 11061D, Thermo Fisher) or CELLection release buffer (CELLection Biotin Binder Kit 11533D, Thermo Fisher) prepared according to the manufacturer’s instructions (release volume of 110 μL).

Techniques: Labeling, Standard Deviation

Journal: ACS Omega

Article Title: Integration of Magnetic Bead-Based Cell Selection into Complex Isolations

doi: 10.1021/acsomega.7b01427

Figure Lengend Snippet: Cell viability following capture and release. (A) The viability of cells (LNCaP, 22Rv1) bound to nonreleaseable beads compared to untouched cells (underwent no magnetic bead isolation) following a 3-day culture (abbreviations: SM = Sera-Mag). (B) The viability of cells bound to (bound) and released from (released) releasable beads (CELLection, FlowComp) following a 3-day culture. Bars represent the average of three technical replicates; error bars represent the standard deviation.

Article Snippet: For experiments involving release, a similar experimental design was followed, except that once washed, the beads and bound cells were dropped into a release well containing either FlowComp Release Buffer (FlowComp Flexi Kit 11061D, Thermo Fisher) or CELLection release buffer (CELLection Biotin Binder Kit 11533D, Thermo Fisher) prepared according to the manufacturer’s instructions (release volume of 110 μL).

Techniques: Isolation, Standard Deviation

Journal: ACS Omega

Article Title: Integration of Magnetic Bead-Based Cell Selection into Complex Isolations

doi: 10.1021/acsomega.7b01427

Figure Lengend Snippet: Characterization of nucleic acid extraction with cell isolation magnetic beads present. (A,B) Relative fold change in the mRNA transcript (HPRT) detected from LNCaPs. Isolations containing cell isolation beads were compared to no-bead controls for two methods of RNA extraction: (A) spin columns and (B) bead-based extraction. (C,D) Similarly, relative fold change in GAPDH from DNA extracted via (C) spins columns or (D) bead-based extraction. Bars represent the average of three technical replicates; error bars represent the standard deviation; * denotes p < 0.05 and *** denotes p < 0.001; --- indicates the cell only, no-bead control (abbreviations: CELL = CELLection, SM = Sera-Mag, FC = FlowComp).

Article Snippet: For experiments involving release, a similar experimental design was followed, except that once washed, the beads and bound cells were dropped into a release well containing either FlowComp Release Buffer (FlowComp Flexi Kit 11061D, Thermo Fisher) or CELLection release buffer (CELLection Biotin Binder Kit 11533D, Thermo Fisher) prepared according to the manufacturer’s instructions (release volume of 110 μL).

Techniques: Extraction, Cell Isolation, Magnetic Beads, RNA Extraction, Standard Deviation, Control