Chamberlin was born in New York City. She was the daughter of Ida Roberta (née Small; 1909-1993) and Bernando LaPallo (died 2015), author of Age Less/Live More and a centenarian.

Her first role in film was a small part in Up the Sandbox starring Barbra Streisand. She had a prominent role as Madame Zenobia in the film Uptown Saturday Night and the follow-up Let's Do it Again. She also appeared in several television films including Long Journey Back (1978), Brave New World (1980), and Once Upon A Family (1980). Her final film role was in the short film Habeaus Corpus (2013).

Chamberlin was a regular performer during the first two years of the esteemed series The Electric Company, and she made guest appearances in the television series What's Happening!!, Diff'rent Strokes, and NYPD Blue. In 1979, she played the wife of James Earl Jones's character on the short-lived police drama Paris. Most notably she played Odile Harris in Roots: The Next Generations (1979). Her first recurring role in a major television sitcom was as Lucy Daniels in "All's Fair" from 1976-77. In the 1970s she appeared on shows such as Lou Grant and James at 16.

In the early 1980s, she appeared on The White Shadow. Other guest spots in the 1980s included Ryan's Four and Beat Street. In 1994; she played Commander Della Thorne in Viper. In 1998, she played Dr. Timmi in The Practice, and Judge Leslie Battles in To Have and To Hold.

Lee began her career in 1968 on in Slave ship, a stage production based on the outline of LeRoi Jones later known as Amiri Baraka. She appeared at The Orpheum Theatre in a musical production called Do Your Own Thing, based on Shakespeare's Twelfth Night, and in an off-Broadway production, The Believers. She played Cordelia opposite James Earl Jones's King Lear in 1974 in the Delacorte Theatre at the New York Shakespeare in the Park Festival. She went on to win six AUDELCO Awards for Excellence in Black Theater on November 21, 1988, for her musical play Struttin’, performed at the Rosetta LeNoire AMAS Repertory Theater. She also appeared in the play Hospice produced at The Henry Street Settlement Theatre in Lower Manhattan.

In 1999, she made guest appearances on Moesha and NewsRadio as Mrs Leveaux. In 2000 she appeared in City of Angels and Any Day Now as Mrs. Samuels. From 1982-90, Chamberlin played Pat Baxter, the mother of Angela Baxter Hubbard on All My Children. In 1997, she appeared in Diagnosis Murder (television series) as Judge Gwen Mosford. In 2002, she appeared on episodes of Touched by An Angel and Judging Amy.

She wrote and acted in her one-woman play Objects in the Mirror are Closer than They Seem first as a reading in Miami, Florida, and later in 2010 as part of The Kitchen Theatre's Counter series in Ithaca, New York from February 10-14 in a sold out run. The play was directed by Rachel Lampert. Chamberlin founded a non-profit organization, Lee Chamberlin's Playwrights' Inn Project Inc., establishing it in France to nurture the work of African American playwrights.

Chamberlin died of cancer at the age of 76 on May 25, 2014 in Chapel Hill, North Carolina.