The Physical Nature of Mesoscopic Inhomogeneities in Highly Diluted Aqueous Suspensions of Protein Particles

Abstract

The dispersed composition of water samples prepared by the multiple-dilution technique has been investigated opto-physically using two laser diagnostics methods: dynamic light scattering and laser phase microscopy. We studied suspensions of antibodies (Ab) to the human interferon-gamma (IFNγ) in water with an initial concentration of 0.125 mg ml⁻¹ (5×10¹⁴ mol cm⁻³), which were subjected to 12 successive 100-multiple volume dilutions (C12) in water. Individual antibodies and particles of about 300 nm in size, which can be considered as aggregates of individual antibodies, have been found in the initial suspensions. It turned out that some mesoparticles with a concentration on the order of 10³ cm⁻³ exist also in suspensions subjected to C12 dilution. These particles have a refractive index close to typical values of protein refractive indices. A possible explanation of the origin of particles with sizes on the order of several hundreds of nanometers, revealed in highly diluted suspensions, is proposed. On the one hand, some of these particles may be aggregates of antibodies from the initial suspension, which remained (due to the flotation effect) in the bulk of the liquid after dilutions. On the other hand, the appearance of solid contaminant particles at the same mesoscopic scale (i.e., on the order of several hundreds of nanometers) cannot be excluded in the dilution process.