Maleimide Crosslinkers

Maleimide Crosslinkers

Published on 21.03.2023

Maleimides provide versatile options for connecting payloads to proteins and other biomolecules. Discover how our bifunctional thiol reactive crosslinkers can support your project. Read on!

Maleimide Crosslinkers

Maleimides readily react with thiols in a Michael type addition to form succinimidyl thioethers. This reaction may be carried out in buffered aqueous solutions at pH 6.8 (useful range: 6.5-7.0) and usually completes within a few minutes. However, this reaction and linkage is reversible.

Schematic representation of the thioether formation between a thiol and a maleimide.

If increased stability of the conjugate is required, the resulting succinimide ring may be deliberately hydrolyzed. To this end, we are providing self-stabilizing crosslinkers with Boc-protected amines, which, after deprotection, may act as intramolecular nucleophiles to open the succinimide ring.

Hydrolysis of thiosuccinimides via an intrinsic catalytic nucleophilic amine. P = Payload.

Besides, we offer linkers with aryl moieties, which directly destabilize the succinimide after it has been formed, so that the ring then is quickly and irreversibly hydrolyzed.

Resonance promoted hydrolysis of thiosuccinimides. P = Payload.

Within our maleimido-crosslinker portfolio, we offer several kinds of additional reactive functions with various properties: reactivity towards amino groups, for use in Click Chemistry, for photoactivation, or to introduce new functional groups into your structure or biomolecule.

In the following, we are highlighting those additional functionalizations:

Succinimidyl esters (abbreviated as OSu or NHS) are a frequently used tool for coupling small molecules to peptides and proteins. These activated carboxyl groups react readily with amino groups in buffered aqueous solutions at pH 7.5 (useful range: 6.5-8.5). Iris Biotech is offering a wide selection of ready to use maleimide-succinimide crosslinkers with spacers of different properties and lengths between 6 and 16 atoms.

Pentafluorophenyl (PFP-) esters work like succinimidyl esters but are slightly less sensitive to undesired hydrolysis during the coupling process. As an example, Mal-L-Dap(Boc)-OPfp (MAA1080) introduces a spacer of five atoms.

Regarding Click Chemistry, we offer DBCO (dibenzoazacyclooctyne) and MeTz (methyltetrazine) functionalized maleimide crosslinkers suitable for copper-free conjugation in biological environments, as well as alkyne-functionalized ones for classical Click chemistry (see related products).

As photoactivatable crosslinker, we provide 4-(N-Maleimido)benzophenone (LS-3350), which can be activated upon irradiation with light of 350 nm.

Further functional groups may be introduced using maleimide-crosslinkers bearing amino, hydroxyl and carboxyl functions.

Besides “permanent” crosslinkers, our portfolio also includes cleavable linkers with a predetermined breaking point for targeted payload release, e.g. upon enzymatic activity, changes in pH or redox potential.

→ Browse our Brochure Linkerology® to discover the panoply of available linker technologies.
→ For more details about Click Chemistry or Photochemistry, download our booklets!
→ You cannot find what you are looking for? Get in contact for a Custom Synthesis and benefit of our expertise!

References:

Reactivity and pH dependence of thiol conjugation to N-ethylmaleimide: detection of a conformational change in chalcone isomerase; R. A. Bednar; Biochemistry 1990; 29: 3684-3690. https://doi.org/10.1021/bi00467a014

Stabilization of cysteine-linked antibody drug conjugates with N-aryl maleimides; R. J. Christie, R. Fleming, B. Bezabeh, R. Woods, S. Mao, J. Harper, A. Joseph, Q. Wang, Z. Q. Xu, H. Wu, C. Gao,N. Dimasi; J Control Release 2015; 220: 660-70. https://doi.org/10.1016/j.jconrel.2015.09.032

A versatile acid-labile linker for antibody–drug conjugates; M. C. Finniss, K. S. Chu, C. J. Bowerman, J. C. Luft, Z. A. Haroon,J. M. DeSimone; Med. Chem. Commun. 2014; 5: 1355-1358. https://doi.org/10.1039/c4md00150h

Long-term stabilization of maleimide-thiol conjugates; S. D. Fontaine, R. Reid, L. Robinson, G. W. Ashley,D. V. Santi; Bioconjug Chem 2015; 26: 145-52. https://doi.org/10.1021/bc5005262

Bioconjugation - A versatile and essential research tool; G. Hermanson; Aldrichimica Acta 2017; 50: 43-57.

Chapter 3 - The Reactions of Bioconjugation; G. T. Hermanson; in Bioconjugate Techniques (Third Edition); edited by G. T. Hermanson; Academic Press; Boston 2013: 229-258 https://doi.org/10.1016/B978-0-12-382239-0.00003-0

Cross-linking of rabbit skeletal muscle troponin with the photoactive reagent 4-maleimidobenzophenone: identification of residues in troponin I that are close to cysteine-98 of troponin C; J. Leszyk, J. H. Collins, P. C. Leavis,T. Tao; Biochemistry 1987; 26: 7042-7. https://doi.org/10.1021/bi00396a028

Self-hydrolyzing maleimides improve the stability and pharmacological properties of antibody-drug conjugates; R. P. Lyon, J. R. Setter, T. D. Bovee, S. O. Doronina, J. H. Hunter, M. E. Anderson, C. L. Balasubramanian, S. M. Duniho, C. I. Leiske, F. Li,P. D. Senter; Nature biotechnology 2014; 32: 1059-62. https://doi.org/10.1038/nbt.2968

pH Dependence of Succinimide-Ester-Based Protein Cross-Linking for Structural Mass Spectrometry Applications; E. Trabjerg, A. Keller,A. Leitner; ACS Measurement Science Au 2022; 2: 132-138. https://doi.org/10.1021/acsmeasuresciau.1c00032

Novel Silyl Ether-Based Acid-Cleavable Antibody-MMAE Conjugates with Appropriate Stability and Efficacy; Y. Wang, S. Fan, D. Xiao, F. Xie, W. Li, W. Zhong,X. Zhou; Cancers (Basel) 2019; 11: 957. https://doi.org/10.3390/cancers11070957

Discovery of Peptidomimetic Antibody-Drug Conjugate Linkers with Enhanced Protease Specificity; B. Wei, J. Gunzner-Toste, H. Yao, T. Wang, J. Wang, Z. Xu, J. Chen, J. Wai, J. Nonomiya, S. P. Tsai, J. Chuh, K. R. Kozak, Y. Liu, S. F. Yu, J. Lau, G. Li, G. D. Phillips, D. Leipold, A. Kamath, D. Su, K. Xu, C. Eigenbrot, S. Steinbacher, R. Ohri, H. Raab, L. R. Staben, G. Zhao, J. A. Flygare, T. H. Pillow, V. Verma, L. A. Masterson, P. W. Howard,B. Safina; J Med Chem 2018; 61: 989-1000. https://doi.org/10.1021/acs.jmedchem.7b01430

Characterization of Ring-Opening Reaction of Succinimide Linkers in ADCs; K. Zheng, Y. Chen, J. Wang, L. Zheng, M. Hutchinson, J. Persson,J. Ji; J Pharm Sci 2019; 108: 133-141. https://doi.org/10.1016/j.xphs.2018.10.063

Verwandte Produkte
    1. Mal-AMCHC-OSu
      Mal-AMCHC-OSu

      Art-Nr.: MAA1000

      Ab 70,00 €

    2. Mal-beta-Ala-OSu
      Mal-beta-Ala-OSu

      Art-Nr.: MAA1020

      Ab 250,00 €

    3. TCEP*HCl
      TCEP*HCl

      Art-Nr.: LS-3405

      Ab 190,00 €

    4. Mal-L-Dap(Boc)-OH*DCHA
      Mal-L-Dap(Boc)-OH*DCHA

      Art-Nr.: MAA1040

      Ab 80,00 €

    5. Mal-D-Dap(Boc)-OH*DCHA
      Mal-D-Dap(Boc)-OH*DCHA

      Art-Nr.: MAA1060

      Ab 90,00 €

    6. Sulfo-SMCC
      Sulfo-SMCC

      Art-Nr.: MAA1050

      Ab 500,00 €

    7. Z-DEVD-Rh110-p
      Z-DEVD-Rh110-p

      Art-Nr.: LS-3550

      Ab 1.210,00 €

    8. MeTz-PEG(4)-mal
      MeTz-PEG(4)-mal

      Art-Nr.: RL-2340

      Ab 260,00 €

    9. Structure image for DBCO-mal
      DBCO-mal

      Art-Nr.: RL-2490

      Ab 240,00 €

    10. Structure image for DBCO-PEG(4)-mal
      DBCO-PEG(4)-mal

      Art-Nr.: RL-2500

      Ab 330,00 €

    11. Structure image for Mal-Hx-NHS
      Mal-Hx-NHS

      Art-Nr.: RL-2660

      Ab 60,00 €

    12. Structure image for Mal-cHxHx-NHS
      Mal-cHxHx-NHS

      Art-Nr.: RL-2650

      Ab 175,00 €

    13. Structure image for 3-Mal-Bz-NHS
      3-Mal-Bz-NHS

      Art-Nr.: RL-2600

      Ab 170,00 €

    14. Structure image for 3-Mal-MBz-NHS
      3-Mal-MBz-NHS

      Art-Nr.: RL-2610

      Ab 240,00 €

    15. Structure image for 4-Mal-Bz-NHS
      4-Mal-Bz-NHS

      Art-Nr.: RL-2620

      Ab auf Anfrage

    16. Structure image for 4-Mal-MBz-NHS
      4-Mal-MBz-NHS

      Art-Nr.: RL-2630

      Ab 220,00 €

    17. Structure image for Mal-Bu-NHS
      Mal-Bu-NHS

      Art-Nr.: RL-2640

      Ab 100,00 €

    18. Structure image for Mal-Pen-NHS
      Mal-Pen-NHS

      Art-Nr.: RL-2670

      Ab 180,00 €

    19. Structure image for Mal-PhBu-NHS
      Mal-PhBu-NHS

      Art-Nr.: RL-2680

      Ab 600,00 €

    20. Structure image for Mal-PrHx-NHS
      Mal-PrHx-NHS

      Art-Nr.: RL-2690

      Ab 260,00 €

    21. DBCO-PEG(24)-MAL
      DBCO-PEG(24)-MAL

      Art-Nr.: PEG6780

      Ab 400,00 €

    22. Mal-NH2*HCl
      Mal-NH2*HCl

      Art-Nr.: RL-2780

      Ab 75,00 €

    23. Mal-Et-OH
      Mal-Et-OH

      Art-Nr.: RL-3000

      Ab 275,00 €

    24. Structure image for Mal-L-Dap(Boc)-OPfp
      Mal-L-Dap(Boc)-OPfp

      Art-Nr.: MAA1080

      Ab 95,00 €

    25. Mal-cyclobutane-1,1-dicarboxamide-Cit-PAB
      Mal-cyclobutane-1,1-dicarboxamide-Cit-PAB

      Art-Nr.: ADC1460

      Ab auf Anfrage

    26. Mal-cyclobutane-1,1-dicarboxamide-Ala-PAB
      Mal-cyclobutane-1,1-dicarboxamide-Ala-PAB

      Art-Nr.: ADC1560

      Ab auf Anfrage

    27. Structure image for Mal-CH2CH2-N(Me)-CH2-COOH
      Mal-CH2CH2-N(Me)-CH2-COOH

      Art-Nr.: RL-3400

      Ab auf Anfrage

    28. Structure image for Mal-AMCHC-N-Propargylamide
      Mal-AMCHC-N-Propargylamide

      Art-Nr.: MAA1100

      Ab 120,00 €

    29. MC-Val-Ala-PAB-Cl
      MC-Val-Ala-PAB-Cl

      Art-Nr.: ADC1700

      Ab 500,00 €

    30. Mal-N-Boc-Aeg-NHS
      Mal-N-Boc-Aeg-NHS

      Art-Nr.: RL-3430

      Ab 195,00 €