Chamberlayne is a surname. Notable people with the surname include:
- Edward Chamberlayne (1616–1703), English writer
- Tankerville Chamberlayne (1840–1924), English Member of Parliament
- Thomas Chamberlayne (disambiguation) several men with this name
- William Chamberlayne (poet) (1619–1679), English poet
- William Chamberlayne (MP) (1760–1829), English Member of Parliament and landowner
- The Chamberlayne family, an influential family of Weston, Southampton, United Kingdom
- The Chamberlayne family, an influential family in Virginia, descended from William Chamberlayne (burgess) of Hereford who settled in New Kent County, Colony of Virginia
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