SNAP-Tag Technology: A Promising Tool for Ex Vivo Immunophenotyping.
SNAP-tag, a self-labeling protein tag, is commonly used for in vitro and in vivo analysis of bound target proteins. We report the first evidence that SNAP-tag could be used for ex vivo detection of enriched biological markers.



Proof of concept was established for target c-kit receptor, a pathological and diagnostic marker for a variety of cancers. SNAP-tag conjugates with stem-cell factor (SCF) fusion proteins were designed and their binding and specificity was validated in vitro using flow cytometry and immunostaining.



Ex vivo diagnostic application of the fusion protein was demonstrated in comparison with anti-c-kit antibody for peripheral blood samples from leukemia patients and colorectal tissue specimens.
Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai, 600036, India.
Swati
S
Choudhary
Department of Pharmaceutical Product Development, Fraunhofer Institute for Molecular Biology and Applied Ecology IME, 52074, Aachen, Germany. stefan.barth@uct.ac.za.|UCT Faculty of Health Sciences, South African Research Chair in Cancer Biotechnology, Cape Town, South Africa. stefan.barth@uct.ac.za.
Stefan
S
Barth
Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai, 600036, India. vermars@iitm.ac.in.
Rama S
RS
Verma
D002460:Cell Line; D015179:Colorectal Neoplasms / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D016130:Immunophenotyping / Q000379:methods*; D015470:Leukemia, Myeloid, Acute / Q000276:immunology / Q000378:metabolism*; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000737:chemistry* / Q000235:genetics; D016612:Paraffin Embedding; D019009:Proto-Oncogene Proteins c-kit / Q000032:analysis* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019089:Stem Cell Factor / Q000235:genetics / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D019089:Stem Cell Factor; D019853:O(6)-Methylguanine-DNA Methyltransferase; D019009:Proto-Oncogene Proteins c-kit