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| molecular_weight = 58.4 kD
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'''CRM197'''<ref name="BrökerCostantino2011">{{cite journal|last1=Bröker|first1=Michael|last2=Costantino|first2=Paolo|last3=DeTora|first3=Lisa|last4=McIntosh|first4=E. David|last5=Rappuoli|first5=Rino|author-link5=Rino Rappuoli|title=Biochemical and biological characteristics of cross-reacting material 197 (CRM197), a non-toxic mutant of diphtheria toxin: Use as a conjugation protein in vaccines and other potential clinical applications|journal=Biologicals|volume=39|issue=4|year=2011|pages=195–204|doi=10.1016/j.biologicals.2011.05.004|pmid=21715186}}</ref> is a non-toxic mutant of [[diphtheria toxin]], currently used as a [[carrier protein]] for [[polysaccharides]] and [[haptens]] to make them [[immunogenic]].<ref>{{cite web|title=CRM197 Product Details|url=http://www.reagentproteins.com/product_detail/CRM197_Lyophilized.html|website=Reagentproteins.com|access-date=3 December 2015}}</ref>
There is some dispute about the toxicity of CRM197, with evidence that it is toxic to yeast cells and some mammalian cell lines.<ref>{{cite journal|last1=Qiao|first1=Jian|last2=Ghani|first2=Karim|last3=Caruso|first3=Manual|title=Diphtheria toxin mutant CRM197 is an inhibitor of protein synthesis that induces cellular toxicity|journal=Toxicon|volume=51|issue=3|year=2008|pages=474–477|doi=10.1016/j.toxicon.2007.09.010|pmid= 17988701}}</ref>
 
== Description ==
CRM197 is a genetically detoxified form of diphtheria toxin. A single mutation at position 52, substituting glutamic acid for glycine, causes the ADP-ribosyltransferase activity of the native toxin to be lost. The structural basis for the lack of CRM197 toxicity has recently been elucidated.<ref name="pmid22431623">{{cite journal | vauthors = Malito E, Bursulaya B, Chen C, Lo Surdo P, Picchianti M, Balducci E, Biancucci M, Brock A, Berti F, Bottomley MJ, Nissum M, Costantino P, Rappuoli R, Spraggon G | title = Structural basis for lack of toxicity of the diphtheria toxin mutant CRM197 | journal = Proc Natl Acad Sci U S A | volume = 109 | issue = 14 | pages = 5229–34 | date = April 2012 | pmid = 22431623 | pmc = 3325714 | doi = 10.1073/pnas.1201964109 }}</ref> CRM197 is widely used as a [[carrier protein]] for [[conjugate vaccine]]s. A potential advantage of CRM197 over [[toxoid]]ed proteins is that, because it is genetically detoxified, it retains its full complement of [[lysine]] [[amine]]s for [[Conjugation (organic chemistry)|conjugation]]. There is also evidence suggesting that, compared with tetanus toxoid, there is less carrier-induced suppression of the immune response, especially when there are many individual polysaccharides linked to the same carrier protein.<ref name=":0" /> A summary of the uses and properties of CRM197 has been published.<ref name=":0">{{cite journal | last1 = Shinefield | first1 = H. R. | year = 2010 | title = Overview of the development and current use of CRM(197) conjugate vaccines for pediatric use | journal = Vaccine | volume = 28 | issue = 27| pages = 4335–4339 | doi=10.1016/j.vaccine.2010.04.072 | pmid=20452430}}</ref> CRM197, like diphtheria toxin, is a single polypeptide chain of 535 amino acids (58.4 kD) consisting of two subunits (linked by disulfide bridges).
 
== Manufacturing ==
The gene for CRM197 has been [[Cloning|cloned]] into ''[[Corynebacterium diphtheriae]]'', the bacteria that produces the native toxin.<ref name="pmid6416165">{{cite journal | vauthors = Rappuoli R | title = Isolation and characterization of Corynebacterium diphtheriae nontandem double lysogens hyperproducing CRM197 | journal = Appl Environ Microbiol | volume = 46 | issue = 3 | pages = 560–4 | date = September 1983 | pmid = 6416165 | pmc = 239316 | doi = 10.1128/aem.46.3.560-564.1983 }}</ref> Like the wild type toxin, CRM197 is expressed as a secreted protein at relatively low yields (typically <100&nbsp;mg/L). Corynebacterium expressed CRM197 is available from several sources, including [[List Laboratories]] and [[SigmaAldrich]]. The low yield and high cost of commercially available native CRM197 has led to efforts to produce CRM197 in other bacteria but this has proven a difficult task until recently.
 
Three companies have succeeded at manufacturing CRM197 as a recombinant protein. Ligand's wholly-owned subsidiary, [https://www.crm197.com/ Pelican (previously Pfenex)], a [[San Diego]]-based developer of the Pelican Expression Technology™ production platform,<ref>NYSE MKT: PFNX</ref> produces the protein ("PeliCRM™") in India at [[Serum Institute of India]] [[Pune]] using ''[[Pseudomonas fluorescens]]'' and various proprietary expression technologies for high yield. [[Finabio|Fina BioSolutions LLC]] of Rockville, Maryland has achieved multi-gram/L expression of CRM197 in E. coli (“EcoCRMTM”) as an [[intracellular]], properly folded soluble protein. Fina Biosolutions currently provides the protein for pre-clinical use. Recombinant CRM197 is also made in low-mutation Clean Genome® E. coli by [[Scarab Genomics LLC]] where transport of CRM197 into the bacterial cell [[periplasm]] enhances its stability and proper folding.
 
== Uses ==
CRM197 is used as a [[carrier protein]] in a number of approved conjugate vaccines. HibtiterTM, a vaccine to protect against Haemophius influenzae type b, approved by the FDA in 1990, was the first conjugate vaccine to use CRM197 (the vaccine was discontinued in 2007). Pfizer's Prevnar, which in 2000 became the first [[pneumococcal conjugate vaccine]] to gain FDA approval, comprises polysaccharides from pneumococcal [[serotype]]s conjugated to CRM197. A larger number of clinical and pre-clinical conjugate vaccines using CRM197 as the carrier protein are being evaluated. A further example of a vaccine currently in use that is a CRM197 conjugate is the meningitis ACWY vaccine, Menveo, produced by [[GlaxoSmithKline]].<ref>{{Cite web|url=https://www.medicines.org.uk/emc/medicine/27347#companyDetails|title=Menveo Group A, C, W135 and Y conjugate vaccine - Summary of Product Characteristics (SPC) - (eMC)|website=www.medicines.org.uk|access-date=2016-10-01}}</ref> In addition, CRM197 made in the Pelican Expression Technology™ platform is used in Merck's VAXNEUVANCE and Serum Institute's Pneumosil.
 
CRM197 possess a binding site for EGF receptor heparin-binding epidermal growth factor-like growth factor (HB-EGF), a member of the EGF family.<ref name="pmid7836353">{{cite journal | vauthors = Mitamura T, Higashiyama S, Taniguchi N, Klagsbrun M, Mekada E | title = Diphtheria toxin binds to the epidermal growth factor (EGF)-like domain of human heparin-binding EGF-like growth factor/diphtheria toxin receptor and inhibits specifically its mitogenic activity | journal = J Biol Chem | volume = 270 | issue = 3 | pages = 1015–9 | date = January 1995 | pmid = 7836353 | doi = 10.1074/jbc.270.3.1015 | doi-access = free }}</ref> As this receptor is overexpressed on cancer cells, there have been efforts to use CRM197 as an anti-cancer therapy.<ref>{{cite journal | vauthors = Buzzi S, Rubboli D, Buzzi G, Buzzi AM, Morisi C, Pironi F | title = CRM197 (nontoxic diphtheria toxin): effects on advanced cancer patients | journal = Cancer Immunol Immunother | volume = 53 | issue = 11 | pages = 1041–8 | date = November 2004 | pmid = 15168087 | doi = 10.1007/s00262-004-0546-4 }}{{cite journal | vauthors = Dateoka S, Ohnishi Y, Kakudo K | title = Effects of CRM197, a specific inhibitor of HB-EGF, in oral cancer | journal = Med Mol Morphol | volume = 45 | issue = 2 | pages = 91–7 | date = June 2012 | pmid = 22718294 | doi = 10.1007/s00795-011-0543-6 }}</ref> The cancer immunotherapy company Imugene reported dramatic improvements in antibody titers from its B cell peptide cancer immunotherapy targeting HER2 when it used CRM197 as a carrier protein.[5]
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Preclinical studies have shown that CRM197 is also suitable for conjugation and presentation of peptide [[epitopes]], a vaccinal approach that could have applications in [[Streptococcal infection]],<ref name="pmid23249976">{{cite journal |vauthors=Caro-Aguilar I, Ottinger E, Hepler RW, Nahas DD, Wu C, Good MF, Batzloff M, Joyce JG, Heinrichs JH, Skinner JM| title = Immunogenicity in mice and non-human primates of the Group A Streptococcal J8 peptide vaccine candidate conjugated to CRM197 | journal = Hum Vaccin Immunother | volume = 9 | issue = 3 | pages = 488–96 | date = Dec 2012 | pmid = 23249976 | doi=10.4161/hv.23224 | pmc=3891704}}</ref> cancer,<ref name="pmid28183282">{{cite journal |vauthors=Tobias J, Jasinska J, Baier K, Kundi M, Ede N, Zielinski C, Wiedermann U| title = Enhanced and long term immunogenicity of a Her-2/neu multi-epitope vaccine conjugated to the carrier CRM197 in conjunction with the adjuvant Montanide | journal = BMC Cancer | volume = 17 | issue = 1 | pages = 118 | date = Feb 2017 | pmid = 28183282 | doi=10.1186/s12885-017-3098-7 | pmc=5301331}}</ref> or [[Alzheimer's disease]]<ref name="pmid27900387">{{cite journal |vauthors=Vingtdeux V, Zhao H, Chandakkar P, Acker CM, Davies P, Marambaud P| title = A modification-specific peptide-based immunization approach using CRM197 carrier protein: Development of a selective vaccine against pyroglutamate Aβ peptides | journal = Mol Med | volume = 2016 | pages = 841–849 | date = Nov 2016 | pmid = 27900387 | doi=10.2119/molmed.2016.00218 | pmc=5263057}}</ref> therapy.
 
== History ==
In 1971 [[Tsuyoshi Uchida]], in the laboratory of [[Alwin Max Pappenheimer Jr.|Alwin Pappenheimer]] at Harvard, used [[nitroguanidine]] to create mutants of diphtheria toxin, which were called Cross Reacting Materials, or CRMs.<ref>{{cite journal
|author1=Uchida T |author2=Gill DM |author3=Pappenheimer AM |title=Mutation in the Structural Gene for Diphtheria Toxin carried by Temperate Phage β.
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