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Chamberlain studied at the Union Theological Seminary, New York (1857-1859). He was ordained by the Presbyterian Church in the United States of America (Presbytery of Rio de Janeiro) in July 8, 1866.
Chamberlain moved to Brazil for medical reasons. He was having eye problems and his doctor advised him to take a nautical trip in order to reestablish his vision to normal conditions. He arrived at Rio de Janeiro in July 21, 1862. He brought with him a recommendation letter for Alexander Latimer Blackford, who was already working in Brazil by them. Chamberlain visited São Paulo and Rio Grande do Sul, Brazilian provinces where he worked as English teacher.
After a short period back in the United States, in order to complete his studies, Chamberlain came back to Brazil where he was ordained by the Presbytery of Rio de Janeiro in 1866.
In 1870, along with his wife, Mary Chamberlain, he started giving classes in his house for protestant children, in what became the Mackenzie Presbyterian University (Portuguese: Universidade Presbiteriana Mackenzie). Chamberlain was one of the main contributors in the development of this institution.
Chamberlain was one of the main leaders of Presbyterianism in Brazil during its first decades of existence.