The Mosbach church in the northern part of Biebrich is the "mother church" of the Biebrich parishes and was first mentioned in 1085. At that time, the main Protestant church in Biebrich belonged to the St. Simeon Abbey in Trier and was probably dedicated to St. Peter the Apostle. In 1472, the Simeon monastery sold the tithes of Mosbach and Biebrich to the Cistercian monastery of Eberbach, which remained responsible for the church in Biebrich and Mosbach until secularization in 1803. It is also the builder of the rectory, the so-called Diltheyhaus and the oldest residential building in Biebrich, as the inscription on the house from 1696 attests. During the Reformation, the church became Protestant in 1560.

The tower of the main Protestant church in Biebrich dates back to the early 16th century. Apart from two late Gothic reliefs of a Holy Sepulchre, nothing of the old church inventory has survived. In addition to a Marian altar, there was also an altar of St. James. It referred to the Brotherhood of St. James that existed here, which looked after pilgrims on their way to Santiago de Compostela. From 1653 to 1802, the main Protestant church in Biebrich also served as a parish church for the Protestants of Mainz.

The main Protestant church in Biebrich took on its current form as a hall building with slightly vaulted barrel ceilings and a rectangular choir in 1712-16. The room is surrounded on three sides by two-storey galleries. During the last renovation (1988-91), the original colors on the columns and the galleries with the Bible verses were uncovered again.

The name Evangelische Hauptkirche only came into use at the beginning of the 20th century, after the Oranier-Gedächtnis-Kirche was built on the banks of the Rhine as another Protestant church.