She has appeared in successful films such as Premam (2015), King Liar (2016), Kavan (2017), Pa Paandi (2017), Iblis (2018), Virus (2019), Brother's Day (2019), Kotigobba 3 (2021), Shyam Singha Roy (2021), Padmini (2023) and Leo (2023).

Madonna was born to Sebastian and Shyla at Cherupuzha, Kerala, India, and she belongs to a Malayali Syrian Christian family raised in Angamaly.

[1] After Premam, she then made her Tamil debut opposite Vijay Sethupathi in Kadhalum Kadandhu Pogum (2016), the film makes mixed to positive reviews from critics, and became an average hit at box office.

[9] In 2018, Madonna Sebastian's first release is a Tamil movie Junga, she landed in an extended cameo appearance in the film, opposite Vijay Sethupathi, marking collaboration with him for the third time,[10] directed by Gokul.

In 2019, her first release was a multi-starrer medical thriller film titled Virus, in which she played the role of Dr. Sara Yakub, directed by Aashiq Abu.

In 2021, Madonna Sebastian made her Kannada debut with Kottigobba 3, opposite Sudeep, directed by Shiva Karthik.

Her second release was Shyam Singha Roy starring Nani and Sai Pallavi, directed by Rahul Sankrityan.

Later she made her digital streaming debut with a Tamil webseries Kaiyum Kalavum, which was released on Sony LIV and received generally positive reviews from critics.

[18] In 2023, her first release was a webseries in Telugu launguage, Anger Tales, which was premiered on Disney + Hotstar and received mixed reviews.

Madonna Sebastian has also sung the jingle composed by Deepak Dev for Hit 96.7, a radio station based in Dubai.