A practical approach to crosslinking

Abstract

The various aspects of chemical crosslinking are addressed. Crosslinker reactivity, specificity, spacer arm length and solubility characteristics are detailed. Considerations for choosing one of these crosslinkers for a particular application are given as well as reaction conditions and practical tips for use of each category of crosslinkers.

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Abbreviations

ABH:

azidobenzoyl hydrazide

ANB- NOS:

N-5-azido-2-nitrobenzoyloxysuccinimide

ASIB:

1-(p-azidosalicylamido)-4-(iodoacetamido)butane

ASBA:

4-(p-azidosalicylamido)butylamine

APDP:

N-[4-(p-azidosalicylamido) butyl]-3′(2′-pyridyldithio)propionamide

APG:

p-azidophenyl glyoxal monohydrate

BASED:

bis-[β-(4-azidosalicylamido)ethyl] disulfide

BMH:

bismaleimidohexane

BS3 :

bis(sulfosuccinimidyl) suberate

BSOCOES:

bis[2-(succinimidooxycarbonyloxy)ethyl]sulfone

DCC:

N,N′-dicyclohexylcarbodiimide

DFDNB:

1,5-difluoro-2,4-dinitrobenzene

DMA:

dimethyl adipimidate·2HCl

DMP:

dimethyl pimelimidate·2HCl

DMS:

dimethyl suberimidate·2HCl

DPDPB:

1,4-di-(3′,2′-pyridyldithio)propionamido butane

DMF:

dimethylformamide

DMSO:

dimethylsulfoxide

DSG:

disuccinimidyl glutarate

DSP:

dithiobis(succinimidylpropionate)

DSS:

disuccinimidyl suberate

DST:

disuccinimidyl tartarate

DTSSP:

3,3′-dithiobis (sulfosuccinimidylpropionate)

DTBP:

dimethyl 3,3′-dithiobispropionimidate·2HCl

EDC or EDAC:

1-ethyl-3-(3-dimethylaminopropyl)carbodimide hydrochloride

EDTA:

ethylenediaminetetraacetic acid disodium salt, dihydrate

EGS:

ethylene glycolbis(succinimidylsuccinate)

GMBS:

N-γ-maleimidobutyryloxysuccinimide ester

HSAB:

N-hydroxysuccinimidyl-4-azidobenzoate

HEPES:

4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

MBS:

m-maleimidobenzoyl-N-hydroxysuccinimide ester

MES:

4-morpholineethanesulfonic acid

NHS:

N-hydroxysuccinimide

NHS-ASA:

N-hydroxysuccinimidyl-4-azidosalicylic acid

PMFS:

phenylmethylsulfonyl fluoride

PNP-DTP:

p-nitrophenyl-2-diazo-3,3,3-trifluoropropionate

SAED:

sulfosuccinimidyl 2-(7-azido-4-methylcoumarin-3-acetamide) ethyl-1,3′-dithiopropionate

SADP:

N-succinimdyl (4-azidophenyl)1,3′-dithiopropionate

SAND:

sulfosuccinimidyl 2-(m-azido-o-nitrobenzamido)-ethyl-1,3′-dithiopropionate

SANPAH:

N-succinimidyl-6(4′-azido-2′-nitrophenyl-amino)hexanoate

SASD:

sulfosuccinimidyl 2-(p-azidosalicylamido)ethyl-1,3′-dithiopropionate

SATA:

N-succinimidyl-S-acetylthioacetate

SDBP:

N-hydroxysuccinimidyl-2,3-dibromopropionate

SIAB:

N-succinimidyl(4-iodoacetyl)aminobenzoate

SMCC:

succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate

SMPB:

succinimidyl 4-(p-maleimidophenyl) butyrate

SMPT:

4-succinimidyloxycarbonyl-α-methyl-α-(2-pyridyldithio)-toluene

sulfo-BSOCOES:

bis[2-sulfosuccinimidooxycarbonyloxy) ethyl]sulfone

sulfo-DST:

disulfosuccinimidyl tartarate

sulfo-EGS:

ethylene glycolbis(sulfosuccinimidylsuccinate)

sulfo-GMBS:

N-γ-maleimidobutyryloxysulfosuccinimide ester

sulfo-MBS:

m-maleimidobenzoyl-N-hydroxysulfosuccinimide ester

sulfo-SADP:

sulfosuccinimidyl(4-azidophenyldithio)propionate

sulfo-SAMCA:

sulfosuccinimidyl 7-azido-4-methylcoumarin-3-acetate

sulfo-SANPAH:

sulfosuccinimidyl 6-(4′-azido-2′-nitrophenylamino)hexanoate

sulfo-SIAB:

sulfosuccinimidyl(4-iodoacetyl)aminobenzoate

sulfo-SMPB:

sulfo-succinimidyl 4-(p-maleimidophenyl)butyrate

sulfo-SMCC:

sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate

SPDP:

N-succinimidyl 3-(2-pyridyldithio)propionate

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Mattson, G., Conklin, E., Desai, S. et al. A practical approach to crosslinking. Mol Biol Rep 17, 167–183 (1993). https://doi.org/10.1007/BF00986726

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Key words

  • carbodiimide
  • crosslinkers
  • homobifunctional
  • heterobifunctional
  • imido esters
  • maleimide
  • NHS esters
  • phenyl azide
  • pyridyl disulfide