His father was Thomas A. Biddle (1776–1857), the son of American Revolutionary War soldier Clement Biddle (1740–1814). His mother was the former Christine Williams (1780–1861). Their fourth child, he was educated in Pennsylvania, enrolling at the University of Pennsylvania in 1834 and graduating in 1838. In 1842, he joined his father's company, where he worked until the beginning of the Civil War.
In 1849, he joined the first troop of the Philadelphia City Cavalry in which he served until 1858. In 1855 he married Julia Williams Rush (1833–1898) daughter of Samuel Rush (1795–1859) and granddaughter of Benjamin Rush. On September 1, 1862, he joined the 121st Pennsylvania Volunteer Infantry, serving under his cousin Chapman Biddle, with whom he fought at the Battle of Fredericksburg, the Battle of Chancellorsville, the Battle of Gettysburg (under Abner Doubleday) and the Battle of Bristoe Station.
After the war he retired from Thomas Biddle & Company, and in 1874 he was named a director of the Pennsylvania Railroad. He also served as a director for the Philadelphia Savings Fund Company and the Lehigh Navigation Company among others, and as executor of the estate of James Rush (1786–1869), was instrumental in the erection of the Ridgway Library (part of the Library Company of Philadelphia).
- Alexander Williams Biddle was born July 4, 1856, married Anne McKennan on 1879, had five children, and died on September 16, 1916.
- Henry Rush Biddle was born March 15, 1858 and died January 2, 1877.
- Julia Rush Biddle was born July 25, 1859 and died February 24, 1885.
- James Wilmer Biddle was born November 22, 1861, married first Cora Rowland, then Elizabeth Southall Clarke, and died November 22, 1927.
- Louis Alexander Biddle was born March 12, 1863, married Rene H. A. Docobu and died in Vichy, France during World War II on October 1, 1940.
- Mariamne Biddle was born November 8, 1866 and died March 4, 1917.
- Lynford Biddle was born August 26, 1871 and died January 24, 1941.
- "Biddle family papers". University of Delaware Library. Retrieved March 23, 2011.
- Meghan Vacca (June 2006). "Biddle Family Papers" (PDF). Historical Society of Pennsylvania. Archived (PDF) from the original on 29 March 2011. Retrieved March 23, 2011.
- Kerry Webb (June 1997). "Union Order of Battle: Army of the Potomac". Gettysburg Discussion Group. Retrieved March 23, 2011.
- John Heiser. "Union Order of Battle: Army of the Potomac". National Park Service. Retrieved March 23, 2011.