Tyrosine Conjugation Methods for Protein Labelling

Álvarez-Dorta, Dimitri; Deniaud, David; Mével, Mathieu; Gouin, Sébastien G.

doi:10.1002/chem.202001992

Cited by 62 publications

(47 citation statements)

References 105 publications

(167 reference statements)

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“…In the preparation of this manuscript, a complementary review on tyrosine modification was published and we wanted to explicitly acknowledge the excellent reviewing in the area by Gouin and co-workers. 11…”

Section: Introductionmentioning

confidence: 99%

Tyrosine bioconjugation – an emergent alternative

et al. 2020

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Section: Introductionmentioning

confidence: 99%

Tyrosine bioconjugation – an emergent alternative

et al. 2020

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“…Of interest, even if only cysteine is labeled we find that a considerable subset of proteins remained consistently identifiable, although labeling additional residue types does increase accuracy, the number of identifiable proteins, and robustness against labeling errors. It should also be noted that the set of residue types targeted for FRET X fingerprinting can be expanded even further; labeling of, e.g., methionine ( Lin et al., 2017 ) or tyrosine ( Alvarez Dorta et al., 2020 ) may be employed to further increase accuracy or tailor our method to the detection of a given target protein.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of FRET X for single-molecule protein fingerprinting

et al. 2021

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Summary Single-molecule protein identification is an unrealized concept with potentially ground-breaking applications in biological research. We propose a method called FRET X (Förster Resonance Energy Transfer via DNA eXchange) fingerprinting, in which the FRET efficiency is read out between exchangeable dyes on protein-bound DNA docking strands and accumulated FRET efficiencies constitute the fingerprint for a protein. To evaluate the feasibility of this approach, we simulated fingerprints for hundreds of proteins using a coarse-grained lattice model and experimentally demonstrated FRET X fingerprinting on model peptides. Measured fingerprints are in agreement with our simulations, corroborating the validity of our modeling approach. In a simulated complex mixture of >300 human proteins of which only cysteines, lysines, and arginines were labeled, a support vector machine was able to identify constituents with 95% accuracy. We anticipate that our FRET X fingerprinting approach will form the basis of an analysis tool for targeted proteomics.

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“…Therefore, in future experiments, it is essential to make use of more site-specific targeting approaches (his-tag coupling, sortase A, etc.) (33)(34)(35) to avoid interfering with the binding affinity of the biomolecule.…”

Section: Discussionmentioning

confidence: 99%

Radiolabeling of Human Serum Albumin With Terbium-161 Using Mild Conditions and Evaluation of in vivo Stability

et al. 2021

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Targeted radionuclide therapy (TRNT) is a promising approach for cancer therapy. Terbium has four medically interesting isotopes (149Tb, 152Tb, 155Tb and 161Tb) which span the entire radiopharmaceutical space (TRNT, PET and SPECT imaging). Since the same element is used, accessing the various diagnostic or therapeutic properties without changing radiochemical procedures and pharmacokinetic properties is advantageous. The use of (heat-sensitive) biomolecules as vector molecule with high affinity and selectivity for a certain molecular target is promising. However, mild radiolabeling conditions are required to prevent thermal degradation of the biomolecule. Herein, we report the evaluation of potential bifunctional chelators for Tb-labeling of heat-sensitive biomolecules using human serum albumin (HSA) to assess the in vivo stability of the constructs. p-SCN-Bn-CHX-A”-DTPA, p-SCN-Bn-DOTA, p-NCS-Bz-DOTA-GA and p-SCN-3p-C-NETA were conjugated to HSA via a lysine coupling method. All HSA-constructs were labeled with [161Tb]TbCl3 at 40°C with radiochemical yields higher than 98%. The radiolabeled constructs were stable in human serum up to 24 h at 37°C. 161Tb-HSA-constructs were injected in mice to evaluate their in vivo stability. Increasing bone accumulation as a function of time was observed for [161Tb]TbCl3 and [161Tb]Tb-DTPA-CHX-A”-Bn-HSA, while negligible bone uptake was observed with the DOTA, DOTA-GA and NETA variants over a 7-day period. The results indicate that the p-SCN-Bn-DOTA, p-NCS-Bz-DOTA-GA and p-SCN-3p-C-NETA are suitable bifunctional ligands for Tb-based radiopharmaceuticals, allowing for high yield radiolabeling in mild conditions.

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Tyrosine Conjugation Methods for Protein Labelling

Cited by 62 publications

References 105 publications

Tyrosine bioconjugation – an emergent alternative

Tyrosine bioconjugation – an emergent alternative

Evaluation of FRET X for single-molecule protein fingerprinting

Radiolabeling of Human Serum Albumin With Terbium-161 Using Mild Conditions and Evaluation of in vivo Stability

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