In an impressive demonstration of both technologies, scientists at Confocal.nl stained SK-BR-3 cells with Sandia Biotech's anti-Her2-RFP FluorAbody. Sandia Biotech’s FluorAbodies are unique probes that provide a consistent and cost-effective option to monoclonal antibodies. The molecular architecture of a FluorAbody, consisting of a fluorescent protein linked to an ScFv, results in an intrinsically fluorescent probe with a 1:1 fluorophore to epitope ratio. These distinctive features allow for true quantitation of biomarkers, prevent cross-linking of their targets, and shorten staining protocols by making blocking or secondary antibodies superfluous. Combined, these properties tackle many shortcomings of conjugated, monoclonal antibodies.

Trastuzumab (Herceptin™) is a monoclonal antibody (huMAb4D5-8) that targets the Her2 receptor protein and is widely used in immunotherapy of breast cancer.

4D5-8 single-chain variable fragment (scFv) was used to construct the Anti-Her2-RFP FluorAbody.

Importance: 1:1 Ratio of binding site to Fluorescent Protein

Better quantitation and reproducible reagent performance compared to fluorochrome-labeled antibodies

Human Breast Cancer Cell Line SK-BR-3; ErbB2 Staining with Anti-Her2 RFP FluorAbody®

SK-BR-3 cells were fixed with 4% paraformaldehyde containing DAPI and incubated with anti-Her2-mRFP. Nuclei are blue and ErbB2 (as detected by anti-Her2-RFP) is labeling the cellular membrane (red). Cells were imaged in epifluorescence with TRITC and DAPI filters and a 63x oil objective.

Formalin Fixed and Parafin-Embedded Tissue; Human ErbB2 Staining Shows High Her2 Positive Breast Cancer Tissue

Immunofluorescence staining and detection of Her2 (ErbB2) biomarker in formalin-fixed and parafin-embedded breast tissue from a Her2 positive tissue sample. This result clearly shows that the Anti-Her2 FluorAbody can be used as a diagnostic reagent in breast cancer tissue.

Triple Negative Epithelial Human Breast Cancer Cell Line MDAMB231 Labeled with Anti-EphA2-Cerlulean FluorAbody®

Immunofluorescence analysis of the anti-EphA2-cerulean biomarker. (A) Labeling of the EphA2 receptor with primary and secondary antibody fluorochrome labeled with AF488. (B) Labeling of EphA2 with the anti-EphA2-cerulean FluorAbody. The epithelial human breast cancer cell line, MDAMB231, was stimulated with dimeric ephrinA1-F. The AF488 labeled sample shows a more diffuse staining pattern when compared to the distinct punctate staining pattern shown using the anti-EphA2- cerulean FluorAbody. This result clearly shows that the FluorAbody reagent is superior and provides a more specific staining pattern with more biologically relevant imaging results.

Sandia Biotech Products

FluorAbodies

Anti-Her2 RFP FluorAbody

SuperFolder GFP

SuperFolder GFP Fluorescence in Thermophilus

Fold N Glow

N-terminus linked ferritin folds correctly and C-terminus folds poorly

Cut N Glow

Bind N Glow

Sandia Biotech Application Notes

Application Note 1

Application Note 2

Sandia Biotech is proud to announce a strategic partnership with Confocal.nl, the Dutch Microscopy Company. Please join us as we combine our FluorAbody® technology with a game-changing confocal imaging platform that provides super-resolution capability with higher sensitivity than most confocal microscopes. Confocal.nl has advanced the art for super resolution and demonstrated low laser power (see confocal.nl challenge) that enables important biologically relevant applications using spheroids, organoids, and tumoroids. Sandia Biotech and Confocal.nl in partnership with Spirochrome, a Swiss Company, are advancing the art of live cell imaging with no wash, multicolor fluorogenic probes for live cell fluorescence nanoscopy. https://bright-i.nl/home-7/the-challenge/

Timelapse of approximately 1 hour of HUVECs stained for Actin-RFP (red), Mitochondria-GFP (green), and Nucleus-SiR DNA (magenta). Imaged with the Re-scan Confocal Microscope (RCM) on an Olympus IX83 with 1.5NA 100x TIRF objective. Laser power for GFP: 1 microwatt, RFP: 1 microwatt, SiR DNA: 3 microwatts. Sample Courtesy: Philippa Phelp, Amsterdam UMC, location VUmc, Amsterdam.