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This study analyzed English language learner performance (ELL) on the June 2012 Biology Massachusetts Comprehensive Assessment System (MCAS). This study examined ELL performance on the overall instrument and across content domains. It further examined the impact of English proficiency. The results indicated that English proficiency was a strong predictor of performance and that ELLs at higher levels of English proficiency overwhelmingly passed. The results further indicated that English proficiency introduced a construct-irrelevant variance on the Biology MCAS and raised validity issues for using this assessment at lower levels of English proficiency.