Custom Peptide Synthesis
AP PEPTIDE provides high-quality peptide synthesis service with a 95% peptide synthesis success rate, which is well above the industry standard of 75%. We are committed to Total Quality Management (TQM) to ensure our customers’ complete satisfaction. TQM includes MS and HPLC analyses performed after the completion of peptide synthesis, purification, and QC (quality control) steps. These analyses are followed by independent QA (quality assurance) procedures, which double guarantee the highest quality possible for every delivered peptide.
Peptide Synthesis Capacity and Capability
Our custom peptides are synthesized with peptide synthesizers, HPLCs, and mass spectrometers capable of producing over 10,000 peptides per month. Other features of our capacity and capability include:
- Capacity: 10,000 peptides / month.
- Flexible quantities: From mg to kg.
- Flexible purities: Crude, desalt, and from 70% to 98% purity.
- Synthesis technologies: Combination of solid phase and liquid phase synthesis, and microwave and ligation technologies
- Modifications: Include phosphorylation, methylation, acetylation, amidation, stable isotope labeling, and many more.
- Quality control: MS and HPLC analyses after each synthesis step, plus additional QA procedures for every peptide.
Range of Services:
Our staffs are experienced in the synthesis of high quality peptides with various modifications. With over 200 units of analytical and preparative HPLC, we can provide peptides from small scale (mgs) to large scale (kgs) in very fast delivery:
- Simple to complex and Linear to cyclized
- Long sequences peptide up to 140 mer
- From small to large scale
- Different purity level (desalted, >75%, >85%, >90%, >95%, >98% and even above 99%)
- Widest variety of modifications (from biotinylation to phosphorylation, to dye labeling, and much more…)
- Peptides – Protein Bioconjugation
- Purification at different scales
- Highest success rate in the industry
High-Quality Peptide Synthesis Platform
Custom synthesized peptides from AP PEPTIDE are manufactured under strict high-level quality control processes. Our ISO 9001:2008certified, Total Quality Management (TQM) platform ensures that each custom peptide is triple checked for quality via both mass spectrometry (MS) and high performance liquid chromatography (HPLC) analyses after each step during peptide purification and quality control (QC) procedures. As the final step in our TQM platform, we perform additional quality assurance (QA) procedures for every custom peptide to further guarantee the delivery of high-quality peptides.
Even after multiple rounds of purification, small amounts of impurities may still exist in the final peptide product. Therefore, AP PEPTIDE uses reversed-phase high performance liquid chromatography (RP-HPLC) to analyze the purity of each custom peptide.
What is RP-HPLC?
RP-HPLC is a widely used analytical tool, in which the components of a complex mixture can be efficiently separated. The analyte mixture is usually dissolved in water, which is sometimes also mixed with an organic solvent or an acid to assist dissolution prior to RP-HPLC analysis. The analyte is carried by a mobile phase consisting of water and an organic modifier, which is pumped through a column packed with a stationary phase which usually is a number of small diameter particles consisting of carbon chains of a specified length on the surface. As the mixture is pumped through the column, the analytes (in this case your custom peptide and small impurities) adsorb to the hydrophobic surface of the stationary phase. As the percentage of organic modifier in the mobile phase is gradually increased, the analytes desorb into the mobile phase. Desorption of a specific analyte is based on its intrinsic properties, thus each analyte will remain in the column for a specific amount of time, called the retention time. Since peptide bonds maximally absorb UV light at the wavelength of 220 nm, a UV spectrometer is commonly used in RP-HPLC to detect a peptide as it elutes from the column. The detection signal is converted to a visual graph called a chromatogram, which is the plot of UV absorbance vs. elution time.
High resolution of eluted peptides is mainly based on the selection of columns and the organic modifier, especially its elution gradient. The height of a theoretical plate of each column used in the manufacture is regularly checked, and different gradients are established to separate varying peptides. Both processes are aimed at maintaining high resolution of every custom peptide. AP PEPTIDE takes special care in developing RP-HPLC protocols to ensure the most effective separation and the accurate purity determination of your custom peptides.
We use electrospray ionization mass spectrometry (ESI-MS) analysis to confirm the molecular weight of the target peptide.
What is ESI-MS?
ESI-MS is an analytical tool used to study macromolecules, where the mass-to-charge ratios (m/z) of all ionized analytes are respectively determined. As one of the widely used soft ionization techniques, electrospray ionization produces a whole charged ion from the solution of analytes without fragmentation of the macromolecule, which provides a reliable and accurate way to measure the molecular weight of the macromolecule. Generally peptides can be ionized into multiple-charged states, either positively or negatively, depending on the number of basic and acidic residues in the sequence.
The ionized peptides are then transferred into a mass analyzer such as a quadruple or a time-of-flight (TOF), which can separate all charged ions according to their m/z. All these isolated ions are respectively detected by an ion detector such as an electron multiplier and finally are converted into a visual graph called a mass spectrum, which is the plot of ion abundance vs. m/z value of all detected ions. The mass spectrum displays all detected m/z signals and the relative intensity of the ions, which is relative to the highest m/z peak. The highest peak is set as 100%. For most peptides, the mass spectrum displays more than one m/z peak representing a series of charge states. Theoretically the m/z values of these peaks should all result in the same calculated molecular mass, however, calculations from these peaks actually vary with each other slightly due to limited resolution of the instrument. In our MS report, the calculated molecular weight from the most abundant m/z peak of the analyte is utilized to largely eliminate variations from the instrument. AP PEPTIDE performs regular cleaning and calibration of mass spectrometers to ensure the mass accuracy and reproducibility of MS analyses.
List of Modification
|N,C-terminal Modification||Special Amino acid||Fluorescence/Dye labeling||Cyclic Peptide||Quenched Fluorescent Peptide||Multiple Antigenic Peptide System||Protein Conjugation|
|Acetylation (N-Terminal),HYNIC (N-Terminal),
Fatty acid (N-Terminal)
Myristic acid (N-Terminal)
Succinylation (Suc, N-Terminal)
|D-Arg, D-Cys, D-Asp, D-Asn, D-Glu, D-Gln, D-Ser, D-His, D-Thr, D-Trp, D-Leu,D-Ile, D-Met, D-Pro, D-Val ,D-Phe, pGlu, Hyp, D-Lys,D-Tyr, D-Orn, Orn, Abu, Aib, (D)1-Nal, (D)2-Pal, (D)4-Cl-Phe, Nva, Nle, Hse, Hcy, Pen, Mpa,N-Methyl amino acid (Ala, Phe, Leu, Ile, Val, Gly, Met),Other amino acid, Dinitrobenzoylation (Lys), Lys(Me2), Phosphorylation (Tyr, Ser, Thr, single site), Tyr(SO3H2), Ser(octanoic acid)
|Biotin (N-Terminal, Y/N Ahx),Biotin (Lys in sequence)
Biotin (without Lys in sequence)
FITC/5-FAM (N-Terminal, Y/N Ahx)
Dansyl (N-Terminal, Y/N Ahx)
|Disulfide bridge 1stDisulfide bridge 2nd
Disulfide bridge 3rd
Amide cyclic (Side chain, end)
|Abz/ Tyr (3-NO2),EDANS/DABCYL||Asymmetric 4 branchesAsymmetric 8 branches||KLHBSA|
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Peptide_Sequence (from N-terminal to C-terminal):
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