Sealing of a microfluidic cartridge is a challenge, because the cartridge commonly contains heat-sensitive biomolecules that must also be protected from contamination. In addition, the objective is usually to obtain a sealing method suitable for mass production. Laser welding is a rapid technique that can be accomplished with low unit costs. Even though the technique has been widely adopted in industry, the literature on its use in microfluidic applications is not large. This paper is the first to report the effects of laser welding on the performance of the heterogeneous immunoassay in a polystyrene microfluidic cartridge in which biomolecules are immobilized into the reaction surface of the cartridge before sealing. The paper compares the immunoassay performance of microfluidic cartridges that are sealed either with an adhesive tape or by use of laser transmission welding. The model analyte used is thyroid stimulating hormone (TSH). The results show that the concentration curves in the laser-welded cartridges are very close to the curves in the taped cartridges. This indicates, first, that laser welding does not cause any significant reduction in immunoassay performance, and second, that the polystyrene cover does not have significant effect on the signal levels. Interestingly, the coefficients of variance between parallel samples were lower in the laser-welded cartridges than in the taped cartridges.