Chamberlain's started his college career at the University of Missouri, where he made the All-Big Eight Conference freshman team. Chamberlain transferred to Wayne State College, a Division II College in Nebraska where he broke most of the receiving records. Chamberlain was a standout wide receiver at Wayne State from 1993-94. In his two seasons with the Wildcats, Chamberlain had 161 catches for 1,941 yards and 14 touchdowns and became the school's all-time leading receiver, In 1993, Chamberlain had 78 catches for 1,015 yards and seven touchdowns to help the 9-1 Wildcats lead NCAA Division II in total offense that season. He added another 83 catches for 926 yards in 1994. The 83 catches tied most in a single season by a Wildcat receiver and his 16 catches vs. Bemidji State that season ranked most in a single game by a WSC receiver. His quarterback was Brett Salisbury, both were roommates and eventual Harlon Hill Trophy finalists. Chamberlain and Salisbury are the only two players to ever be nominated in the school's history for the Harlon Hill Award.
Chamberlain was drafted by the Denver Broncos in the 7th round (222nd overall) of the 1995 NFL Draft. He played in the NFL for nine seasons. Byron played for Denver Broncos, Minnesota Vikings, and the Washington Redskins. He spent the 1996 World League season with the Rhein Fire and led the league with 58 receptions for 685 yards (11.8) and a league leading 8 touchdowns. Byron was a key contributor as a member of the Denver Broncos back to back Super Bowl wins (Super Bowl XXXII & XXXIII). He was voted to the 2002 Pro Bowl while with the Minnesota Vikings. Byron finished his career with 167 catches for 2,048 yards (12.3) with 6 touchdowns.
In the fall of 2008, Chamberlain returned to Wayne State in Wayne, Nebraska to complete his degree. In an afternoon ceremony, Friday December 19, 2008 Chamberlain received his Bachelor's degree in Communications after nine seasons in the NFL.
Chamberlain started the Byron Chamberlain Foundation in 2000 to assist underserved individuals and communities. Some of the foundations programs have included sports camps for underprivileged kids, clothes and shoe drives, food drives, toy drives as well as health education. Byron looks forward to adding new programs to his foundation including college scholarships as well other health education initiatives