This study examined the relationship between handedness, second-to-fourth digit ratio (2D:4D), and personality traits among the Ikwerre people of Rivers State, Nigeria. The aim was to determine whether biological factors such as digit ratio and hand preference are associated with the Big Five personality dimensions, namely Extraversion, Agreeableness, Conscientiousness, Neuroticism, and Openness to Experience. A cross-sectional design was employed, and data were collected using a structured questionnaire and anthropometric measurements. The data were analysed using descriptive statistics, correlation, regression, and analysis of variance. The results revealed that the majority of participants were right-handed, while a small proportion were left-handed or ambidextrous. The mean 2D:4D ratios for both hands were approximately 0.96, which falls within the normal range reported in the literature. Correlation and regression analyses showed no significant relationships between 2D:4D ratios and any of the Big Five personality traits, nor between digit ratio and handedness. The regression model further indicated that handedness and 2D:4D together explained less than one percent of the variance in personality traits. Analysis of variance revealed that handedness had a statistically significant effect on Extraversion (p = 0.017) and a marginal effect on Openness to Experience (p = 0.066), with left-handed individuals showing slightly higher scores in these traits. However, no significant associations were found for Agreeableness, Conscientiousness, or Neuroticism. In conclusion, the findings suggest that biological markers such as 2D:4D ratio and handedness have minimal influence on personality traits in this population, indicating that environmental and socio-cultural factors may play a more dominant role in shaping personality.