The goal of the paper is to present the preparation, characterization and testing of a new type of polyelectrolyte based on acrylamide and hydrolyzed collagen for flocculation purposes. Two kinds of hydrolyzed collagen with different molecular weights have been obtained from tanned leather by-products. Col I type having a molecular weight of 14,900 Da was prepared by chemical enzymatic processing and Col II type of 16,300 Da by chemical processing. Polyelectrolytes, also called flocculants, were obtained by electron beam irradiation with doses between 2 and 4.5 kGy in atmospheric conditions and at room temperature. The obtained flocculants were characterized using various physical and chemical methods to determine the conversion coefficient, residual monomer content, intrinsic viscosity, molecular weight and radius of gyration. Also, the flocculants were characterized by Fourier transform infrared spectroscopy technique to highlight the collagen binding to the acrylamide structure and by scanning electronic microscopy to investigate the microstructure. A possible reaction mechanism was proposed. The flocculation characteristics were evaluated in 0.2 wt% kaolin suspension at room temperature.