Low protein adsorption coatings are useful for anti-thrombus coatings and anti-fouling coatings for medical devices, as well as water treatment devices. Layer-by-Layer (LbL) coatings are advantageous in making thin films using water soluble polymers, providing unique and adjustable surface morphologies of polymer thin films, and applying for complicated shape substrates. Here we propose low protein adsorption Layer-by-Layer coatings utilizing anionic polymers with Poly(Ethylene Glycol) (PEG) chain or zwitterionic (ZI) moiety. These polymers are deposited by LbL assembly with cationic polymer on planar substrate. Both PEG and ZI systems showed ultra low protein adsorption properties in both anionic and cationic proteins at physiological conditions. We also studied the effects of LbL assembly conditions such as pH and salt concentration. Those thin films showed lower protein adsorption by adjusting pH and salt concentration. We will discuss the relationship between thin film structures and protein adsorption properties.