The first successful preparation of nanofibers of a polyampholyte (N-carboxyethylchitosan) by electrospinning was achieved by adding a non-ionogenic water-soluble polymer to the spinning solution. Using this approach, other polyelectrolytes, poly(2-acryloylamido-2-methylpropanesulphonic acid) (PAMPS), and copolymers of 2-acryloylamido-2-methylpropanesulphonic acid (AMPS) and acrylic acid [P(AMPS-co-AA)] were also electrospun into nanofibers. The non-ionogenic water-soluble polymers were polyacrylamide (PAAm) and poly(vinyl alcohol) (PVA). The average diameters of the electrospun nanofibers were in the range 50-260 nm. The average diameter of the nanofibers significantly decreased with increasing polyelectrolyte content. The electrospun nanofibers were crosslinked by heat treatment at 100, 120 or 150 degrees C for the N-carboxyethylchitosan/PAAm pair and at 90 degrees C in the case of P(AMPS-co-AA)/PVA. The presence of an ionizable low-molecular-weight compound (7-iodo-8-hydroxyquinoline-5-sulphonic acid, SQ) led to a more than two-fold decrease in the diameter of the nanofibers and to the appearance of defects. The SQ-containing nanofibers showed antimicrobial activity against pathogenic microorganisms.