Primm produces and offers custom synthetic peptides with a wide range of scales, purity levels, modifications and labelings, to allow you high performance in your research.
Over the last 20 years we have built a solid international reputation as a reference company when it comes topeptide synthesis. Peptides play a key role in biochemistry, pharmacology and neurobiology for structural and functional studies. Primm provides the best solution for many applications in all fields of life science.
- Monoclonal antibodies
- Polyclonal antibodies
- Study of antigen-antibody interactions
- Epitope mapping
- Function studies
- Receptor studies
Fmoc-chemistry. The peptide chains are assembled on a solid support starting from the C-terminus and progressing towards the N-terminus, by repeating three basic steps:
- deprotection of the Fmoc alpha amino group of the amino acid
- activation of the next protected amino acid, as an active ester
The SPPS method guarantees a cleaner chemistry process because, while the peptide is being synthesized step by step, all soluble reagents can be removed from the peptide-solid support by filtration and washed away. After the desired sequence of amino acids has been obtained, the peptide is removed from the polymeric support via TFA (Trifluoroacetic Acid) cleavage with concomitant removal of the protecting groups on the amino acid side-chains.
Peptides are purified by Reversed Phase (RP-HPLC). HPLC purification allows the removal of the truncated sequences (deletions), the scavengers and the TFA (Trifluoroacetic Acid) used to deprotect and cleave the peptide from the support (TFA could otherwise be extremely toxic and interfere with the expected activity of the peptide).
Primm offers different purity levels:
- Crude peptides for preliminary experimental screening (e.g. crude peptide libraries)
- > 70% for immunological purposes
- > 95% for biological activity studies (animal studies, cell studies); for biochemical reactions (binding experiments or receptor/ligand studies); for structural studies (NMR, X-ray, MS).
An ultra pure level (> 98%) is also available upon request.
Primm offers a wide range of peptide synthesis scales from 1-2 mg up to hundreds of mg. Larger quantities, yielding up to 1 gram of purified peptides, are also available upon request (bulk synthesis).
Due to the wide-spreading interest in Proteomics, peptide libraries have become an advantageous mean for high throughput screening in studies of protein-protein interaction or peptide/protein cross talking, in antigen and epitope mapping, in protein scanning and characterization and in many other fields.
Primm offers peptide libraries with different features to meet all requirements of your research:
- Size: 24, 48, 96 or multiple peptides (8-15 aa length)
- Purity level: crude, >70%, >80% and >95%
- Format: single tube or 96-well plate
- Lyophilized or resuspended at the desidered volume/concentration
Primm can help the customer in designing the library that best fit his experiment. Usually peptides are 8-15 amino acids long and supplied in 1-2 mg quantity, but different lengths and quantities are also available upon request.
24, 48, 96 or multiple crude peptides – average purity 60-70%
- Gold crude: HPLC and MS spectrum of all peptides
– Delivery: up to 4 weeks
- Silver crude: HPLC of 5 statistically significant peptides and MS spectrum of all peptides – Delivery: 3 weeks
- Bronze crude: HPLC and MS spectrum of 5 statistically significant peptides -Delivery: 3 weeks
Pure Library – >70% >80% >95% –
24, 48, 96 or multiple purified peptides – HPLC and MS spectrum of all peptides.
Delivery: up to 4 weeks, depending on the number of peptides and on the purity level
Storage, Use and Handling
– Biotin NH2-terminal and K-Biotin C-terminal
– 5-6 carboxyfluorescein or 5 carboxyfluorescein NH2-terminal
– 5-6 carboxytetramethyl rhodamine N-terminal
– Cy3/Cy5 N-terminal
– N-terminal acetylation
– C-terminal amidation
– Linkers:Aminohexanoic Acid/8-Amino-3,6-dioxaoctanoic Acid
– Phosphorylation amino acids
– D-amino acids
– Cyclic peptides
– Sepharose-bound peptides
– Conjugated peptides (to carrier protein OVA, KLH or BSA)
– 4 branched MAP resin
Synthetic peptides are normally small molecules, which are not able to elicit a significant immune response. However, when they are coupled to a larger carrier molecule, the immune system will then respond to the hapten-carrier complex producing antibodies against the peptide and the carrier protein. We offer you the choice between the most commonly used carrier-proteins: BSA, KLH, OVA and others upon request.
There are a variety of reagents used for binding the peptide to the carrier:
Glutaraldehyde, which cross-links primary NH2-groups on the peptide to those of the protein.
Sulfo-SMCC (Sulfosuccinimidil 4-N-maleimidomethyl-cyclohexane-1-carboxylate), a heterobifunctional reagent that binds, in a two-step reaction, the primary amines of the carrier protein and the sulphydryl group of the cysteine on the peptide.
For peptide sequences lacking cysteine residues, we suggest the addition of a cysteine either at the N- or C-termini of the peptide depending on several criteria. EDC (1-Ethyl-3-(3 Dimethylaminopropyl) carbodiimide), reacts with a carboxylic group, allowing it to be coupled to the amino group in the reaction mixture.
An alternative system to produce antibodies using peptide-conjugates is to use MAPs (Multiple Antigenic Peptides). In this case, the peptide chain grows on a 4 branching lysine core. Lysine has two amino groups and can be used to build up a large branching lysine complex.
In general, use the 4-branch MAP resin for peptides containing more than 15 amino acid residues and the 8 branch MAP resin for peptides shorter than 15 amino acids.
The desired epitope should be at the distal flexible end of the peptide. This approach produces a higher titer of anti-peptide antibodies, eliminating the presence of contaminant anti-carrier antibodies.
Custom Sythesis of peptides having in their sequence special amino acid derivates bearing stable isotopes ( 2H, 13C, 15N) is also available at Primm. please submit your sequence and enquire for a customized quote.
Please click here to request a quote. Our synthesis specialists will contact you.
How do our prices compare to those of our competitors?
Our prices are quite competitive and in accordance with prices from other Peptide Suppliers. It has to be taken into consideration that the price depends very much on the quality of the final product. The critical manufacturing step, to reach a good quality, is the purification, that is the most expensive and time-consuming step. The more a peptide is difficult, the more the purification will be complicated and, as a conseguence, the more the peptide will be expensive.
Is there any limitation on the length of peptides?
Sequences less than 4 amino acids can be really problematic during the cleavage and the purification. For this reasons, we prefer to evaluate sequences < 5 amino acids. We don’t make dimers. Orders of peptides between 8 up to 25 amino acids in length are always accepted. Primm will also synthesize sequences up to 40 residues and quotes will reflect the quantity and the purity requirements. For longer sequences, please inquiry.
What quantity are we able to produce?
Primm can produce peptides from mg to grams and at different purity levels from crude to 95-98% pure.
What if Primm does not succed in making the requested peptide?
Primm accepts any order for peptides not longer than 25 amino acid residues. If one sequence cannot be made, Primm will inform the customer in approximately 1 week from the order, suggesting eventual modifications to the sequence. Of course, if the sequence cannot be made, any cost will be charged. For sequences longer than 25 amino acids, Primm staff must see them before accepting the order and the acceptance will be communicated within 24 hours. Again, if Primm won’t be able to make the peptide, the customer won’t be requested to pay any fee. To give you a realistic estimation, the synthesis failure at Primm labs, for peptides shorter than 25 aa, is less than 3%.
What purity levels are available and what’s the best purity level for my application?
Primm offers different purity levels starting with > 70% up to > 95%, but it’s also possible an ultra pure level (> 98%) upon request.
Please, refer to the following guideline, to select the most suitable grade of purity for your application:
- non-sensitive screening assays: crude or purity >70%
- immunological applications: purity > 70%
- receptor/ligand studies, bio-assay studies or cell studies (e.g. cellular activation / drug screening in cell culture): purity >95%
- structural studies (NMR, X-ray, MS): purity >95%
If my peptide is 95% pure, what is in the other 5%?
Peptide purity is determined by reverse-phase HPLC. As a limit of peptide synthesis method (= solid phase synthesis), the coupling reaction of one amino acid to another is not always 100% efficient, causing a variety of deletion sequences to be generated. Most of the deletion sequences are purified out, but a few, having similar chromatographic characteristics to the target peptide, remain in the peptide sample and account for the percent of impurities.
How are the peptides supplied and how must be stored?
All peptides are supplied lyophilized, unless differently requested. The lyophilized solids should be stored at -20ï¿½C (or even better at -80ï¿½C) for long time or at +4ï¿½C for short time (< 3 months). When solubilized should be stored at -20ï¿½C / -80ï¿½C for short time (< 3 months).
What Quality Control information is supplied?
With the exception of crude peptides , which will only be checked by MALDI-MASS spectrometry, all synthetic peptides will be analyzed by HPLC and MS to verify composition and purity. The QC documentation is supplied for every peptide and is enclosed to the peptide-vial.
How long does it take from order to shipment?
The manufacturing steps of the peptide-production: synthesis, purification and QC analysis require almost 3 weeks. For very long or hard sequences the production should take up to 4/5 weeks.
What type of chemical modification does Primm provide?
We offer mostly of the commercially available modifications such as: Biotin, Fluorescein, Rhodamin, isotopic amino acids, MAP 2 or 4 branched, Phosphorylated residues, Spacers-Linkers, Cys-Cys or head-tail Cycles. We can supply also other modifications upon request.
How do I dissolve the peptide?
The solubility information is reported on the data sheet enclosed to the peptide. Please, read this carefully before starting. The solubility greatly depends on various parameters (the primary and second structure, the alternation of neutral, hydrophobic and charged amino acids, the nature of the modifications made, the solvent and so on.). Therefore it’s quite impossible to predict the real solubility of the peptide before its actual production.
Please, consider the following suggestions.
As a general role, always begin by reconstituting a small amount of peptide before committing the entire lot. The most common dissolution process is 1 mg of peptide dissolved in 1 mL of sterile water (avoid reconstituting a peptide in a buffer, such as PBS because salts hinder the solubility). If the small amount of peptide is not perfectly solubilized in that conditions (= just water), try the following steps on the left peptide:
- Sonication is helpful to dissolve
- Small amounts of dilute (max 10%) aqueous acetic acid for basic peptides and aqueous ammonia for acidic peptides may help
- Choose the appropriate solvent. Begin reconstituting at a concentration higher than your desired final working concentration (only after the complete dissolution of the peptide, add water or buffer to reach the desired final concentration; note: salts may cause aggregation!)
- H2O: for hydrophilic residues (K-R-H-D-E-N)
- Organic solvents: DMF, DMSO, Acetonitrile or TFA: for hydrophobic residues (A-V-L-I-M-F-W)
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