Peptide synthesis, purification of antibodies and other proteins
These services are provided by Mathias Madalinski and Zsuzsanna Muhari-Portik, located in the IMBA, Plaza floor, room nr. 5.65.1, tel: 3718 or 3829.
Depending on the sequence, peptides can be synthesized up to a length of 35 amino acids. After the synthesis they are routinely purified by reversed phase HPLC. Subsequently, the purified peptides are lyophilized. For "normal" sequences a yield of 10 to 50 mg per peptides can be expected.
We also perform peptide synthesis on a smaller scale in a 96 well format. Here, up to 96 different peptides can be synthesized in parallel. The yield lies in the range of 100 µg to 2 mg and peptides are not HPLC purified. Only if absolutely required purification can be performed for a selected subset of the synthesized peptides.
The principle of the technique is to dispense small droplets of pre-activated amino acid derivatives onto a predefined array of positions on a porous membrane. The droplets get absorbed and form individual reaction compartments for chemical reaction in solid phase synthesis. Up to 600 individual peptides can be synthesized on one Cellulose membrane. (appr 20nmol per spot). SPOT synthesis works best for peptides with a length between 6 and 15 residues and provides a purity of ~80%. Beyond that length the quality drops slowly as not all peptide chains are fully accessibly any more. We offer synthesis of peptides with a maximum length of 25 amino acids. Please find more detailed information here.
Synthesis of peptides carrying modifications or amino acids with heavy isotopes
We also offer synthesis of peptides containing modified amino acids. The following modifications are routinely available in our facility:
- N-term acetylated
- N-term Biotin (optional with aminohexanoic acid spacer)
- N-term 5,(6) Carboxyfluorescein (isomer mix) (optional with aminohexanoic acid spacer)
- C-term -> amid
Besides the above mentioned routine modifications, a variety of other modifications can be synthesized with components provided by the scientist. Some examples are listed below:
- N-term 5 or 6 Carboxyfluorescein (monoisomer) (optional with aminohexanoic acid spacer)
- C-term Biotin (standard with PEG5-Spacer)
- C-term Carboxyfluorescein (monoisomer or isomer mix)
Purification of antibodies, cytokines and enzymes
We perform affinity purification of polyclonal anti-peptide-antibodies using corresponding peptide columns. Additionally, we offer purification of polyclonal and monoclonal antibodies via protein A or protein G columns.
Furthermore, in collaboration with the Molecular Biology Service analytical and preparative purification of recombinant cytokines and enzymes via ion exchange and reversed phase chromatography is provided.
As an additional service we provide KLH-peptide conjugates for immunization of animals.