Peptide synthesis and modification technology services

Experimental Technology Introduction: Peptide synthesis is a solid-phase synthesis sequence that generally starts from the C-terminus (carboxyl end) to the N-terminus (amino end). The past peptide synthesis was carried out in solution, known as liquid-phase synthesis method. Since Merrifield successfully developed the solid-phase peptide synthesis method in 1963, through continuous improvement and *, solid-phase synthesis has become a commonly used technique in peptide and protein synthesis today, demonstrating the advantages of classical liquid-phase synthesis *, greatly reducing the difficulty of product purification in each step. Peptide synthesis is generally divided into two types: solid-phase synthesis and liquid-phase peptide synthesis.

Experimental technology principle:

The peptide synthesizer uses solid phase as the reaction principle, and continuously adds, reacts, and synthesizes amino acids in a known sequence (usually from C-carboxy end to N-terminal amino end) in a sealed explosion-proof glass reactor to obtain the peptide carrier. The solid-phase synthesis method greatly reduces the difficulty of purifying each step of the product. To prevent side reactions, the side chains of the amino acids involved in the reaction are protected. The carboxyl end is free and must be activated before the reaction.

Experimental operation process:

1. Remove protection: The column and monomer protected by Fmoc must be treated with an alkaline solvent (piperidine) to remove the protective group of the amino group.

2. Activation and cross-linking: The carboxyl group of the next amino acid is activated by an activator. Activated monomers react and crosslink with free amino groups to form peptides

The key. In this step, a large amount of high concentration reagents are used to drive the reaction to complete. Cycle: These two steps of reaction are repeated until synthesis is complete.

3. Elution and deprotection: Peptides are eluted from the column, and their protective groups are eluted and deprotected by a deprotection agent (TFA).

Peptide modification type:

1. C-end tagging technology:

C-terminal amidation

Aldehydes, Alcohols, pNA (P-Nitroanilide)

AMC、AFC、 Cystamide, Ester, N-alkylI Amides, etc

2. N-terminal tagging technology:

Acetylated, Palmytoyl, HYNIC, Biotinylated, Bromoacetylated

Chelation reaction (DOTA, DTPA conjugated), formylation, tetradecyl, myristoxylated succinylation, palmitoylation, malic acid, fatty acid, etc. (Succinylated)

3. Fluorescent labeling modification:

C-end modifications: AFC, AMC, Dap (Dnp), Lys (Dye), pNA, Rh110

N端修饰: Bodipy-FL, Cy3, Cy5, Texas Red, 5-Tamra, 5-odoacetamido fluoresceinRhodamine 110 and Rhodamine B Luciferin, EDANS

FAM, FITC, MCA, Rox, Sulforhodamine 101, 5-TAMRA

4. Cyclization reaction:

End to end loop, middle loop, special loop (N ->C or Head to Tail)

2 pairs of disulfide bonds, 3 pairs of disulfide bonds (S-S bond formation). Trulide formation) Natural product active peptides form cyclic natural peptides

5. Methylation modification:

Side chain methylation Lys(For),LysS(Me), Lys(Me)2, Lys(Me)3, Arg(Me)2 symmetrical, D-Tyr(Me),D-Tyr(Et)N End methylation (N-Me Arg, N-Me Asp, N-Me Glu, N-Me Leu, N-Me Nle, N-Me Nova, N-Me Phe, N-Me-S N)-

Me-Ser, N-Me-Trp, N-Me-Thr, N-Me-Val)

6. Several special modifications:

Various conjugates (BSA, KL H conjugated peptides for antibody production), phosphorylation (Phosphateline, Phosphaterenine, Phosphatyrosine), sulfonated tyrosine or serine, MAPS (Multiple Antigenic Peptides), glycopeptides PEG modification (PEGylation)

Experimental precautions:

The customer provides:

1. Peptide sequence, quantity, purity, and special requirements

2. Freeze dried peptides can be stored at -20 degrees Celsius or room temperature

3. The solution peptide is much more unstable than the freeze-dried form, and the solution should be stored at a neutral pH (5-7) -20 degrees Celsius.

4. To avoid repeated freeze-thaw cycles of the sample,haoSeparate into small samples for storage.

Delivery standards:

Taicheng Peptide and Data

Others

The first of most peptidesxuanThe solvent is ultra pure distilled water, and dilute Z-acid or ammonia solution can also help dissolve alkaline or acidic peptides, respectively. If these methods do not dissolve the peptides after use, it is recommended to dissolve them with DMF, urea, guanidiniam, chloride, or acetophenone

ELISA kit free testing and experimental outsourcing quotation