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Protein Expression & Purification Technologies

Protein expression and purification can be very challenging because there is no single approach that is appropriate for every protein or downstream application.

Successful expression can be affected by each individual protein’s solubility, toxicity, need for post-translational modifications or its intrinsic ability to fold. Additionally, the application for which the protein is ultimately to be used may dictate how the protein needs to be expressed or purified. It is therefore important to have a flexible set of tools and methods that can be applied to each unique project.

Each NEB expression technology offers different advantages enabling you to choose the strategy that best suits your protein expression and purification needs. Many of the expression vectors share a compatible polylinker, enabling the gene of interest to be easily shuffled between systems. Various applications along with the recommended NEB kit can be found by clicking the sidebar tab.

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Protein Expression & Purification Technologies includes these subcategories:
Cell-Free Expression
E. coli
Resins, Magnetic Beads & Racks
FAQs for Protein Expression & Purification Technologies
Protocols for Protein Expression & Purification Technologies
Application Notes for Protein Expression & Purification Technologies
    Publications related to Protein Expression & Purification Technologies
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    • Walters, K.J., et al. (2003) DNA -repair protein hHR23a alters its protein structure upon binding proteasomal subunit S5a Proc Natl Acad Sci U S A; 100, 12694-12699.
    • Yee, C.S., et al. (2003) Generation of the R2 subunit of ribonucleotide reductase by intein chemistry: insertion of 3-nitrotyrosine at residue 356 as a probe of the radical initiation process Biochemistry; 42, 14541-14552. PubMedID: 14661967
    • Hong, S.H., and Maret, W. (2003) A fluorescence resonance energy transfer sensor for the beta-domain of metallothionein Proc Natl Acad Sci U S A; 100, 2255-2260. PubMedID: 12618543
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    • Feng, Z., et al. (2011) Optimization of Medium Composition for Production of Recombinant Calf Chymosin from Kluyveromyces lactis in Submerged Fermentation J Northeast Agric Univ; 18, 56-62.
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    • Feng, Z., et al. (2011) Disruption of PMR1 in Kluyveromyces lactis improves secretion of calf prochymosin J Sci Food Agric; 15, 100-103.
    • Narayan, V., Pion, E., Landré, V., Müller, P. and Ball, K.L. (2011) Docking-dependent ubiquitination of the interferon regulatory factor-1 tumor suppressor protein by the ubiquitin ligase CHIP J Biol Chem; 286, 607-619 . PubMedID: 20947504
    • Handa, Y., Inaho, N. and Nameki, N. (2011) YaeJ is a novel ribosome-associated protein inEscherichia coli that can hydrolyze peptidyl-tRNA on stalled ribosomes Nucleic Acids Res; 39, 1739-1748 . PubMedID: 21051357
    • Vernet, E., Kotzsch, A., Voldborg, B., et al. (2011) Screening of genetic parameters for soluble protein expression in Escherichia coli Protein Expr Purif; 77, 104-111. PubMedID: 21130169
    • Sakhtah, H., Behler, J., Ali-Reynolds, A., Causey, T.B., Vainauskas, S., Taron, C.H. (2019) A novel regulated hybrid promoter that permits autoinduction of heterologous protein expression in Kluyveromyces lactos Appl Environ Microbiol; pii: e00542-19. PubMedID: 31053583
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    • Muto, H., Nakatogawa, H., and Ito, K. (2006) Genetically Encoded but Nonpolypeptide Prolyl-tRNA Functions in the A Site for SecM-Mediated Ribosomal Stall Mol Cell; 22, 545-552. PubMedID: 16713584
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    • Villemagne, D., Jackson, R. and Douthwaite, J.A. (2006) Highly efficient ribosome display selection by use of purified components for in vitro translation J Immunol Methods; 313, 140-148. PubMedID: 16730021
    • Hackenberger, C. P., et. al. (2006) Expression of N-terminal Cys-protein fragments using an intein refolding strategy Bioorg Med Chem; 14, 5043-5048.
    • Shimizu, Y. Kanamori, T. and Ueda, T. (2005) Protein synthesus by pure translation systems Methods; 36, 299-304. PubMedID: 16076456
    • Ge, X., et. al. (2005) Self-cleavable stimulus responsive tags for protein purification without chromatography J Am Chem Soc; 127, 11228-9. PubMedID: 16089436
    • Girish, A., et al. (2005) Site-specific immobilization of proteins in a microarray using intein-mediated protein splicing Bioorg Med Chem Lett; 15, 2447-51. PubMedID: 15863295
    • Goodin, J. L., et al. (2005) Yersinia pestis outer membrane type III secretion protein YscC: expression, purification, characterization, and induction of specific antiserum Protein Expr Purif; 40, 152-163. PubMedID: 15721783
    • Colussi, P.A. and Taron, C.H. (2005) Kluyveromyces lactis LAC4 promoter variants that lack function in bacteria but retain full function in K. lactis Appl Environ Microbiol; 71, 7092-7098. PubMedID: 16269745
    • Colussi, P.A., Specht, C.A. and Taron, C.H. (2005) Characterization of a nucleus-encoded chitinase from the yeast Kluyveromyces lactis Appl Environ Microbiol; 71, 2862-2869. PubMedID: 15932978
    • Ying, B.W., Taguchi, H., Kondo, M. and Ueda, T. (2005) Co-translational involvement of the chaperonin GroEL in the folding of newly translated polypeptides J Biol Chem; 280, 12035-12040. PubMedID: 15664980
    • Kuruma, Y., Nishiyama, K., Shimizu, Y., Muller, M and Ueda, T. (2005) Development of a minimal cell-free translation system for the synthesis of presecretory and integral membrance proteins Biootechnol Prog; 21, 1243-1251. PubMedID: 16080708
    • Rebets, Y., et al. (2005) DNA -binding activity of LndI protein and temporal expression of the gene that upregulates landomycin E production in Streptomyces globisporus 1912 Microbiology; 281-290.
    • Morassutti, C., et al. (2005) Expression of SMAP-29 cathelicidin-like peptide in bacterial cells by intein-mediated system Protein Expr Purif; 39, 160-168.
    • Ingham, A.B., et al. (2005) A versatile system for the expression of nonmodified bacteriocins inEscherichia coli J Appl Microbiol; 98, 676-683. PubMedID: 15715871
    • Sando, S., Kanatani, K., Sato, N., Matsumoto, H. Hohsaka, T. and Aoyama, Y. (2005) A small-molecule-based approach to sense codon-templated natural-unnatural hybrid peptides. Selective silencing and reassignment of the sense codon by orthogonal reacylation stalling at the single-codon level J Am Chem Soc; 127, 7998-7999. PubMedID: 15926808
    • Chuzel, L., Ganatra, M.B., Schermerhorn, K.M., Gardner, A.F., Anton, B.P., Taron, C.H. (2017) Complete genome sequence of Kluyveromyces lactis strain GG799, a common yeast host for heterologous protein expression Genome Announc; 5(30), PubMedID: 28751387
    • Feng, Y. and Cronan, J. E. (2009) A new member of the Eschericia coli fad regulon: transcriptional regulation of fadM (ybaW) J Bacteriol; 191, 6320-6328 . PubMedID: 19684132
    • Talabot-Ayer, D., Lamacchia, C., Gabay, C., and Palmer, G. (2009) Interleukin-33 is biologically active independently of Caspase-1 cleavage J Biol Chem; 284, 19420-19426 . PubMedID: 19465481
    • Solaroli, N., Panayiotou, C., Johansson, M., and Karlsson, A. (2009) Identification of two active functional domains of human adenylate kinase 5 FEBS Lett; 583, 2872-2876 . PubMedID: 19647735
    • Wan, X.M. et al. (2009) Identification of nose-to-brain homing peptide through phage display Peptides; 30, 19007831. PubMedID: 343-350
    • Estevez-Torres, A., Crozatier, C., Diguet, A., Hara, T., Saito, H., Yoshikawa, K., and Baigl, D. (2009) Sequence-independent and reversible photocontrol of transcription/expression systems using a photosensitive nucleic acid binder Proc Natl Acad Sci U S A; 106, 12219-12223 . PubMedID: 19617550
    • Zheng, Y., Posfai, J., Morgan, R. D., Vincze, T., and Roberts, R.J. (2009) Using shotgun sequence data to find active restriction enzyme genes Nucleic Acids Res; 37, el . PubMedID: 18988632
    • Chattopadhaya, S., et al. (2009) Use of intein-mediated protein ligation strategies for the fabrication of functional protein arrays Methods Enzymol; 462, 195-223. PubMedID: 19632476
    • Hondal, R. J. (2009) Using chemical approaches to study selenoproteins-Focus on thioredoxin reductases Biochim Biophys Acta; 1790, 1501-1512. PubMedID: 19406205
    • Nothelfer, E.M. et al. (2009) Identification and characterization of a peptide with affinity to head and neck cancer J Nucl Med; 50, 426-434. PubMedID: 19223422
    • Madinger, C.L., Sharma, S.S., Anton, B.P., Fields, L.G., Cushing, M.L., Canovas, J., Taron, C.H. and Benner, J.S. (2009) The effect of carbon source on the secretome of Kluyveromyces lactis Proteomics; 9, 4744-4754. PubMedID: 19743416
    • Mueller, J. et al. (2009) Targeting of tumor blood vessels: a phage-displayed tumor-homing peptide specifically binds to matrix metalloproteinase-2-processed collagen IV and blocks angiogenesis in vivo Mol Cancer Res; 7, 1078-1085. PubMedID: 19584266
    • de Marco, A. (2009) Strategies for successful recombinant expression of disulfide bond-dependent proteins in Escherichia coli Microb Cell Fact; 8, PubMedID: 19442264
    • Che, N., et. al. (2009) Soluble expression and one-step purification of a neurotoxin Huwentoxin-I in Escherichia coli Protein Expr Purif; 65, 154-159. PubMedID: 19217942
    • Chattopadhaya, S., et al. (2009) Site-specific covalent labeling of proteins inside live cells using small molecule probes Bioorg Med Chem; 17, 981-9. PubMedID: 18261914
    • Tanner, D., Cariello, D., Woolstenhulme, C., Broadbent, M. and Buskirk, A. (2009) Genetic identification of nascent peptides that induce ribosome stalling J Biol Chem; 284, 34809-34818 . PubMedID: 19840930
    • Feng, Z., et al (2009) Effect of temperature-shift strategy on recombinat chymosin of Kluyveromyces Lactis Heilongjiang Animal Science and Veterinary Medicine; 2009-2013.
    • Anton, B.P., Fomenkov, A., Raleigh, E.A. and Berkmen, M. (2016) Complete Genome Sequence of the Engineered Escherichia coli SHuffle Strains and Their Wild-Type Parents Genome Announc; Mar 31;4(2), PubMedID: 27034504, DOI: 10.1128/genomeA.00230-16.
    • Dorsch, S., et al. (2001) The VP1-unique region of parvovirus B19: amino acid variability and antigenic stability J Gen Virol; 82 (pt 1) , 191-199 . PubMedID: 11125172
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