Chopra, the third of six children of Ranbir Lal and Rooprani Chopra, a Punjabi family, was born on 23 September 1935, in Lahore. After the partition of India, his family moved to Simla (present-day Shimla), where he was brought up. He has studied in S.D. Sr. Sec. School, Simla. His father desired for him to be a doctor or an Indian Administrative Services Officer.

During his stint with the Times of India, he worked in Punjabi films including Sapani and Hindi films such as Woh Kaun Thi?, Shaheed, Main Shaadi Karne Chala and Teesri Manzil. Prem did not consider acting as a full-time profession in the early 1960s, but he kept trying to get roles in films due to his passion for acting. Among his early films he played Sukhdev in Shaheed, one of his rare positive leading roles. Prem had done four movies before Woh Kaun Thi?, a box office hit of 1964, had released. On the sets of Woh Kaun Thi? which had Manoj Kumar as lead hero, Prem met Manoj for the first time. Manoj offered Prem a positive role in Shaheed, for which Manoj was the presenter. During the shooting of Main Shadi Karne Chala someone suggested that he become a villain. He continued working with The Times of India, when he was already a part of box office hits like Nishan, Sikandar E Azam in 1965 and Sagaai, Mera Saaya in 1966. After Teesri Manzil and Upkaar, he was flooded with films as a villain.

After Upkaar in 1967, he left The Times of India to concentrate solely on acting.

Since 1967, he has been a leading villain in Hindi films and his peak period as main villain was from 1967 to 1995. In the 1970s he got plum roles as villains often with Sujit Kumar and panu Ranjeet. In a few films he played secondary villain to villainous characters of Ajit, Madan Puri, Pran, Prem Nath, Jeevan in the 1970s and 1980s and to Amrish Puri and Amjad Khan in the late 1980s.

In the critically acclaimed comedy film Hulchul (1971), a remake of Anubavam Pudhumai, he played the lead hero in this suspense thriller. He played the main lead in the 1970 box office hit small budget comedy film Samaj Ko Badal Dalo as lead hero, and was paired opposite Telugu film actresses Kanchana and Sharada. The song sung by Rafi and picturised on Prem Chopra – “Tum Apni Saheli Ko Itna Bata Do Ki Usse Koi Pyar Karne Laga Hai” from this 1970 film and the song “Rah Mein Kaliya” – a song sung by Kishore Kumar and performed on-screen by Prem Chopra in the film Nafrat (1973), a box office flop, continues to be popular. The dialogue “Prem naam hai mera, Prem Chopra” (My name is Prem, Prem Chopra) from the film Bobby has been very popular. Another famous Prem Chopra dialogue is from the film Souten – “Main wo bala hoon jo sheshay se pathar ko todte hai”, which closely translates to “I am that trouble which crushes stones with glass”. Another dialogue from Souten was – “Jinke Ghar Sheeshe Ke Hote Hain Woh Batti Bhujakar Kapde Badalte Hai”. “Main jo aag lagata hoon use boojhana bhi jaanta hoon” from Kati Patang.

He was the regular in villainous roles in the films with Rajesh Khanna in the lead role from 1969 to 1991 from Doli (1969) till Ghar Parivar (1991). The duo of Prem and Rajesh Khanna acted in 19 films together and 15 of them were box office hits, and they were very close friends in real life till Khanna’s death. Prem quoted in an interview “Rajesh Khanna and I were considered a lucky pair and distributors used to tell Rajesh Khanna that we don’t care who your heroine is, all we want to know is whether Prem Chopra is in the film.” He also mouthed dialogues in the song “Aaj Pila Dey Saathi Apni” sung by Mahendra Kapoor from the film Doli (1969). He also mouthed dialogues in the song “Lo Mera Pyar Lelo” from Nafrat in 1973 with Asha Bhosle and “Kab Se Ye Dil Hai Pyaasa” from Mera Muqaddar in 1988.

He became estranged with two of his four brothers in the late 1980s. Chopra had bought a bungalow in 1980 in Delhi, which was jointly owned by him and his father, and his father and one brother used to stay there. Chopra had gotten his brother a job in Delhi, and made him stay at the bungalow. But his father, a day before his death, was made to sign a will favouring one of his brothers, taking away Chopra’s rights to the bungalow. Later, an income tax raid happened in that same house, and his brother said in the raid that Chopra had given them the bungalow, but the house still was in name of Prem Chopra. Chopra had two other houses in Bombay as well, which were sold off cheaply by his other brothers without telling him, as they needed the money.

After 1995, he appeared as a villain in very few films. He took up positive character roles from 1996 and started getting more positive roles from 2005. His screen space got reduced in the films after 2005.

His biography titled Prem Naam Hai Mera, Prem Chopra, written by his daughter Rakita Nanda, was released in April 2014.