Poly(tetrahydrofuran) (PTHF) reacted with toluene diisocyanate (TDI) to form a prepolymer. Such a prepolymer was extended by 2,2-hydroxymethyl butanoic acid (DMBA) to form a segmented polyurethane with carboxyl (PUc) consisting of alternating soft and hard segments, where carboxyls are distributed only along the hard segments. By using static and dynamic laser light scattering (LLS) as well as atomic force microscopy (AFM), we have investigated the self-assembly of the PUc and poly(4-vinylpyridine) (P4VP) in tetrahydrofuran/chloroform mixed solvent. Our experiments show that the PUc and P4VP form a stable assembly with a size of 59-122 nm due to the H-bonding, depending upon the PUc (COOH) concentration. The LLS and AFM measurements demonstrate the assembly is spherical.