This study examined the effect of teamwork on organizational performance among selected agro-allied firms in the Sagamu Interchange Business Hub, Ogun State, Nigeria. A survey research design was adopted, and data were collected through structured questionnaires from 235 proportionally selected employees using stratified random sampling. Teamwork was measured through collaboration, communication, and trust, while organizational performance was assessed using productivity, innovation, and operational efficiency. Diagnostic tests confirmed data validity and suitability for regression analysis. The regression results showed that teamwork significantly enhances productivity, with collaboration (β = 0.674, p = 0.000) emerging as the strongest predictor, followed by communication (β = 0.219, p = 0.000) and trust (β = 0.192, p = 0.000). For innovation, collaboration (β = 0.498, p = 0.000) was the only strongly significant factor, while communication was marginally significant (β = 0.134, p = 0.052), and trust was insignificant (p = 0.468). Similarly, operational efficiency was significantly driven by collaboration (β = 0.429, p = 0.000) and communication (β = 0.216, p = 0.001), while trust remained insignificant (p = 0.127). The study concludes that teamwork exerts a significant effect on the performance of employees. It recommends the need for managers in agro-allied firms to foster team collaboration and structured communication systems as strategic levers for enhancing their performance and competitiveness.