Lipidation is a peptide modification that involves the addition of lipid molecule to a peptide sequence. Lipopeptides are of particular interest for vaccine development because it avoids the use of the sometimes toxic adjuvants while still generating a comprehensive immune response. Synthetic peptides alone are not enough to generate an immune response and therefore an immunoadjuvant is typically employed to enhance the response. Alternatives to the classical Freund’s adjuvants have had some success in peptide vaccine work that includes conjugations to bacterial and viral carrier proteins, immunostimulating complexes, and branched multiple antigenic peptides (MAPS).
Lipopeptides first entered into vaccine development after the observation in the early 1980’s by Hopp and co-workers that found improved anti-hepatitis surface antigen response in a peptide conjugated to a dipalmityl-lysine moiety.[1] The reason for this finding is that while short peptides may contain cytotoxic T lymphocytes (CTL) that bind to MHC class I molecules, they fail to prime CD8+ T-cell responses in vivo. This was further confirmed in the early 1990’s by Rammensee and co-workers who showed that an influenza lipopeptide (without the need for an adjuvant) elicited CTL responses whereas the corresponding peptide without lipid conjugation did not.
CPC Scientific has extensive experience and knowledge in synthesizing peptides containing lipid moieties and can link virtually any peptide sequence to lipids that include, palmitoyl, dipalmitoyl, myristic acid, geranyl, farnesyl, geranylgeranyl, and more.
Common Lipids used in SPPS
Common Lipid Abbreviations
| ABBREV. | FULL NAME | IUPAC NAME |
|---|---|---|
| PAL | Palmitic acid | Hexadecanoic acid |
| MYR | Myristic acid | Tetradecanoic acid |
| GER | Geranyl group | (2E)-3,7-dimethylocta-2,6-dien-1-yl |
| FAR | Farnesyl group | (2E,6E)-3,7,11-trimethyldodeca-2,6,10-trien-1-yl |
| GGER | Geranylgeranyl group | (2E,4E,6E)-3,5,7,11-Tetramethyldodeca-2,4,6,10-tetraene-1-yl |
- Hopp, Thomas P. “Immunogenicity of a synthetic HBsAg peptide: enhancement by conjugation to a fatty acid carrier.” Molecular Immunology 21, no. 1 (1984): 13-16.
Peptide Lipidation Citations
Diao, J., Komura, R., Sano, T., Pantua, H., Storek, K.M., Inaba, H., Ogawa, H., Noland, C.L., Peng, Y., Gloor, S.L. and Yan, D. Journal of Bacteriology 203, no. 13 (2021): 10-1128.
- Departments of Infectious Diseases, Structural Biology, Biochemical and Cellular Pharmacology, Translational Immunology, Pathology, Protein Chemistry, Early Discovery Biochemistry, Genentech, South San Francisco, California, USA
For the Lgt biochemical assay, the Pal-IAAC and Pal-IAAA peptides were synthesized by CPC Scientific.
Pantua, H., Skippington, E., Braun, M.G., Noland, C.L., Diao, J., Peng, Y., Gloor, S.L., Yan, D., Kang, J., Katakam, A.K. and Reeder, J. MBio 11, no. 5 (2020): 10-1128.
- Departments of Infectious Diseases, OMNI Bioinformatics, Chemistry, Structural Biology, Biochemical and Cellular Pharmacology, Translational Immunology, Pathology, Molecular Biology, Genentech, South San Francisco, California, USA
MQLNKV-L(U13C6,15N)-KGL(U13C6,15N)MIALPVMAIAA-dipalmitoyl2C-SSNKNGG-K-biotin, which upon cleavage by LspA, yields the product peptide dipalmitoyl2C-SSNKNGG-K-biotin. A product standard [dipalmitoyl2C-SSNKNAAK-(NHCH2CH2NH)-biotin; CPC Scientific] was in the reaction mixture as an internal standard for normalization of product quantitation.
Buss, Colin G., and Sangeeta N. Bhatia. Proceedings of the National Academy of Sciences (2020).
Tandem Peptides. Tandem peptides were purchased from CPC Scientific. Sequences are: mTP-TAMRA-LyP1 (Myr-GWTLNSAGYLLGKINLKALA-ALAKKILGGGGK(5TAMRA)-CGNKRTRGC (C-C bridge)); [..] PEGylated formulations of these peptides were synthesized as previously described (49). The sequences are: [..] mTP-PEG-ARAL (Myr-GWTLNSAGYLLGKINLKALA-ALAKKILC-PEG5K-GGGARALPSQRSR).
Lo, J.H., Hao, L., Muzumdar, M.D., Raghavan, S., Kwon, E.J., Pulver, E.M., Hsu, F., Aguirre, A.J., Wolpin, B.M., Fuchs, C.S. and Hahn, W.C. Molecular Cancer Therapeutics 17, no. 11 (2018): 2377-2388.
pTP-TAMRA-iRGD (CH3(CH)15-[GWTLNSAGYLLGKINLKALAALAKKIL-GGK(TAMRA)GGCRGDKGPDC, Cys-Cys bridge]) used in all figures except Fig. S1 was synthesized by CPC Scientific.
Gilles, Maud-Emmanuelle, Slack, Frank J, et al. Oncotarget, 2019, Vol. 10, (No. 51), pp: 5349-5358
"Tandem peptide (pTP-iRGD: CH3(CH)15-GWTLNSAGYLLGKINLKALAALAKKIL-GGK(TAMRA)GGCRGDKGPDC, Cys-Cys bridge) was synthesized by CPC Scientific."
Kwon, Ester J., et al. Advanced Materials 29.35 (2017).
"[..] synthesized for initial screening with FAM-conjugated lysine at the C-terminal end of the membrane-interactive peptide and with or without D[KLAKLAK]2 on the C-terminus using standard Fmoc chemistry[..] All peptides were synthesized with N-terminal myristic acid and C-terminal amine. [..] larger quantities of the peptides were synthesized by CPC Scientific to 90% purity"
Noland, Cameron L., et al. Proceedings of the National Academy of Sciences 114.30 (2017): E6044-E6053.
- Departments of Structural Biology, Infectious Diseases, Biochemical and Cellular Pharmacology, Pathology, Translational Immunology, and Biomolecular Resource Group, Genentech, Inc., South San Francisco, CA 94080.
"..peptide substrates were synthesized based on the Pal lipoprotein, Pal peptideshort (Pam2Cys-SSNKNGGK-Biotin; CPC Scientific, Inc.).."
Lo, Justin H., et al. Bioconjugate Chemistry (2016).
"Myristoyl-transportan−LyP-1 tandem peptide was synthesized by CPC Scientific (sequence: myr-GGWTLNSAGYLLGKINLKALAALAKKIL-GGGG-CGNKRTRGC, Cys−Cys bridge). FAM-labeled LyP-1 bearing an azide (azidoacetyl-GGG-cyclo-(EGNKRTRGK)), FAM-labeled LyP-1 bearing a cysteine (C-K(5FAM)-C6-cyclo-(CGNKRTRGC)) and transportan bearing a C-terminal cysteine (myristic acid-GGWTLNSAGYLLGKINLKALAALAKKILC) were synthesized by CPC Scientific, Inc."
Kwon, Ester J., Matthew Skalak, Riana Lo Bu, and Sangeeta N. Bhatia. ACS Nano 10, no. 8 (2016): 7926-7933.
"[The peptides] were synthesized by CPC Scientific with N-terminal myristic acid."
Longo, Edoardo, et al. International Journal of Pharmaceutics 472.1 (2014): 156-164.
"[palmitoylated peptides] were synthesized by CPC Scientific Inc. (Sunnyvale, CA, USA), each obtained as a white lyophilised powder and stored at 4 °C protected from light. All peptides were synthesized by peptide solid-phase synthesis."