26.10: The Merrifield Solid-Phase Technique
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Objectives
After completing this section, you should be able to describe, briefly, the Merrifield solid‑phase technique for the synthesis of polypeptides.
Key Terms
Make certain that you can define, and use in context, the key term below.
- solid‑phase method (solid‑phase synthesis)
Study Notes
The solid‑phase used in this method is a polymer support. You will not be examined on the details of the Merrifield solid‑phase method; however, you should be prepared to write a couple of paragraphs describing this important process.
For his work on the synthesis of peptides, Bruce Merrifield was awarded the 1984 Nobel Prize in chemistry.
The synthesis of a peptide of significant length (e.g. ten residues) by this approach requires many steps, and the product must be carefully purified after each step to prevent unwanted cross-reactions. To facilitate the tedious and time consuming purifications, and reduce the material losses that occur in handling, a clever modification of this strategy has been developed. This procedure, known as the Merrifield Synthesis after its inventor R. Bruce Merrifield , involves attaching the C-terminus of the peptide chain to a polymeric solid, usually having the form of very small beads. Separation and purification is simply accomplished by filtering and washing the beads with appropriate solvents. The reagents for the next peptide bond addition are then added, and the purification steps repeated. The entire process can be automated, and peptide synthesis machines based on the Merrifield approach are commercially available. A series of equations illustrating the Merrifield synthesis may be viewed below . The final step, in which the completed peptide is released from the polymer support, is a simple benzyl ester cleavage. This is not shown in the display.
| The Merrifield Peptide Synthesis | |
Two or more moderately sized peptides can be joined together by selective peptide bond formation, provided side-chain functions are protected and do not interfere. In this manner good sized peptides and small proteins may be synthesized in the laboratory. However, even if chemists assemble the primary structure of a natural protein in this or any other fashion, it may not immediately adopt its native secondary, tertiary and quaternary structure. Many factors, such as pH, temperature and inorganic ion concentration influence the conformational coiling of peptide chains. Indeed, scientists are still trying to understand how and why these higher structures are established in living organisms.