Selective immunocapture and light-controlled traceless release of transiently caged proteins

Rasa Rakauskaitė; Giedrė Urbanavičiūtė; Martynas Simanavičius; Aurelija Žvirblienė; Saulius Klimašauskas

doi:10.1016/j.xpro.2021.100455

Selective immunocapture and light-controlled traceless release of transiently caged proteins

STAR Protoc. 2021 Apr 14;2(2):100455. doi: 10.1016/j.xpro.2021.100455. eCollection 2021 Jun 18.

Authors

Rasa Rakauskaitė¹, Giedrė Urbanavičiūtė¹, Martynas Simanavičius¹, Aurelija Žvirblienė¹, Saulius Klimašauskas¹

Affiliation

¹ Institute of Biotechnology, Life Sciences Center, Vilnius University, Vilnius LT-10257, Lithuania.

Abstract

The 4,5-dimethoxy-2-nitrobenzyl (DMNB) photocaging group introduced into small biomolecules, peptides, oligonucleotides, and proteins is commonly used for spatiotemporal control of chemical and biological processes. Here, we describe the use of a DMNB-selective monoclonal antibody for non-covalent capture of chemically or biosynthetically produced proteins containing surface-exposed DMNB caging groups followed by light-controlled traceless decaging and release of the bound proteins into solution for a variety of downstream applications. For complete details on the use and execution of this protocol, please refer to Rakauskaitė et al. (2020).

Keywords: Antibody; Molecular/Chemical Probes; Protein Biochemistry.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antibodies / chemistry*
Fluoresceins / chemistry*
Light*
Peptides / chemistry*

Substances

Antibodies
Fluoresceins
Peptides