Slight differences in the amino acid sequence of a protein. For example, to produce a recombinant protein in E. coli, a methionine (met) must be added to one end of the protein sequence to act as a signal that initiates protein synthesis. In most cases, that met is removed once the protein is made. Sometimes the met is removed for only some of the molecules. The purified product is then a mixture of a protein with the native sequence plus the extra amino acid.