More than 600 members of the United Methodist Church (UMC) say Attorney General Jeff Sessions' endorsement of a new "zero tolerance" immigration policy infringes upon church law.
In the complaint, the 640 signatories accuse Mr Sessions of "child abuse" by introducing "practices that indefinitely separate thousands of young children from their parents" and "holding thousands of children in mass incarceration facilities with little to no structured educational or socio-emotional support."
Taking a tougher stance on people illegally entering the country from Mexico has led to 2,000 children being separated from their families at the border, recent official statistics revealed.
Last week, Mr Sessions defended the new immigration policy by quoting chapter 13 of Romans, where Paul urges the believers in Rome to respect the God-given authority of their earthly leaders.
In response, the complaint accuses him of spreading "doctrines contrary to the standards of doctrine of the United Methodist Church", through the "misuse of Romans 13 to indicate the necessity of obedience to secular law."
Among the other charges clergy and lay members levelled against Mr Sessions - who is a UMC member and attends UMC churches in Alabama and Virginia - are "immorality" and "racial discrimination".
Premier has contacted The United States Department of Justice for a comment.
Amid widespread criticism after footage of children being held in cages and crying was release, President Donald Trump defended the policy and said illegal migrants were "infesting" the country.
The formal complaint could technically lead to a church trial; however, this is thought to be unlikely.
Rev William Lawrence, an expert on the UMC, was quoted by CNN as saying such a move would be "highly unusual".
The complaint has been championed by Rev David Wright, chaplain at the University of Puget Sound in Tacoma, Washington.
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