Journal: iScience
Article Title: Affimer reagents enable targeted delivery of therapeutic agents and RNA via virus-like particles
doi: 10.1016/j.isci.2024.110461
Figure Lengend Snippet: Impact of Affimers on the metabolic activity of breast cancer cell lines A range of breast cancer cell lines AU-565, SKBR3, BT-474, MDA-MB-453, MCF-7, MDA-MB-231, and ZR-75-1 were seeded one day prior to the addition of HER2-binding Affimers or HER2-binding Affimer-MMAE conjugates. Cells were incubated for a further 72 h, and metabolic activity was analyzed by AlamarBlue measurement. HER-2-binding Affimers did not affect metabolic activity in either HER2-positive cell lines AU-565 and BT474 or the HER-2-negative cell line MDA-MB-231 [(A) and (B)]. HER2-binding Affimers were conjugated with the cytotoxin MMAE via a cathepsin cleavable group and a PABC spacer (C). Seven breast cancer cell lines with varied HER2 expression level as measured by immunoblotting (D); representative blot shown; dotted line indicates removal of a lane containing lysates from a non-breast cancer cell line were treated with the Affimer-MMAE conjugates and showed dose-dependent inhibition of metabolic activity [(E) and (F)], the efficacy of which varied with HER2 expression level. All values were normalized to cells incubated with media alone. Data are mean ± SEM; dose-response curves were fitted using GraphPad Prism v 9.0, [Inhibitor] vs. response -- Variable slope (four parameters); n = 3 independent experiments for all panels. MMAE, monomethyl auristatin E.
Article Snippet: HER-2 ECD-FC tagged , Sino Biologicals , Cat#10004-H04H.
Techniques: Activity Assay, Binding Assay, Incubation, Expressing, Western Blot, Inhibition
![Impact of Affimers on the metabolic activity of breast cancer cell lines A range of breast cancer cell lines AU-565, SKBR3, BT-474, MDA-MB-453, MCF-7, MDA-MB-231, and ZR-75-1 were seeded one day prior to the addition of HER2-binding Affimers or HER2-binding Affimer-MMAE conjugates. Cells were incubated for a further 72 h, and metabolic activity was analyzed by AlamarBlue measurement. <t>HER-2-binding</t> Affimers did not affect metabolic activity in either HER2-positive cell lines AU-565 and BT474 or the HER-2-negative cell line MDA-MB-231 [(A) and (B)]. HER2-binding Affimers were conjugated with the cytotoxin MMAE via a cathepsin cleavable group and a PABC spacer (C). Seven breast cancer cell lines with varied HER2 expression level as measured by immunoblotting (D); representative blot shown; dotted line indicates removal of a lane containing lysates from a non-breast cancer cell line were treated with the Affimer-MMAE conjugates and showed dose-dependent inhibition of metabolic activity [(E) and (F)], the efficacy of which varied with HER2 expression level. All values were normalized to cells incubated with media alone. Data are mean ± SEM; dose-response curves were fitted using GraphPad Prism v 9.0, [Inhibitor] vs. response -- Variable slope (four parameters); n = 3 independent experiments for all panels. MMAE, monomethyl auristatin E.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_8639/pmc11298639/pmc11298639__gr3.jpg)
