Simultaneous separate and group determination of tylosin and tilmicosin in foodstuffs using single antibody-based immunoassay

► Rabbit polyclonal antibody was produced against tylosin (TYL)-bovine serum albumin conjugate. ► Changes in coating antigen design could modify the specificity of indirect competitive ELISAs. ► TYL-selective and equal for TYL and tilmicosin (TMN) recognition were reached. ► Convenient test for simultaneous and separate TYL/TMN determination was developed. ► Screening examination of milk, eggs, honey and chicken muscle was carried out.