After studying Local Governance Administration in the North-West Academy of Public Administration in St. Petersburg, from which he graduated in 2006, he completed a correspondence course at the Saint Tikhon's Orthodox University in Moscow.
His political career began in 1991, when he joined the Free Democratic Party of Russia. From 1994 to 1995, he was an assistant to Vitaly Viktorovich Savitsky, chairman in the 1990s of the existing Christian Democratic Union of Russia (CDU). During this time Milonov was also chairman of the "Young Christian Democrats", a political youth branch of the CDU Russia. Later he became a friend of the Russian politician Galina Starovoitova who supported him politically as well. After the murder of Starovoitova, he turned away from politics. In 2004 he began again his political career at the municipal level as a council member of the community "Dachnoe", in 2005 as head of the city administration of Krasnenkaya Rechka Municipal Okrug in St. Petersburg. In 2007 he successfully ran for election to the Legislative Assembly of Saint Petersburg and was in his first term Chairman of the Committee for the establishment of the government, local government and territorial management structure. In 2009, he moved to the Chair of the Committee for legislation. In 2011 he was re-elected as an MLA. While in the legislature, Milonov was the principal sponsor of legislation criminalizing "homosexual propaganda directed toward minors". In 2016 elections, he was elected to the State Duma representing United Russia.
He is married and has two children. In 1991 he joined the Baptist church. Later, in 1998, he converted to the Russian Orthodox Church.
In 2013, Milonov stated that gay athletes could be subject to arrest at the Sochi Olympics, if promoting homosexuality to minors. He stated, "If a law has been approved by the federal legislature and signed by the president, then the government has no right to suspend it. It doesn't have the authority." Milonov also protested the Side by Side LGBT film festival in November 2013.
On October 30, 2014, Milonov spoke out about Apple CEO Tim Cook's homosexuality and said on the FlashNord website: “What could he bring us? The Ebola virus, AIDS, gonorrhea? They all have unseemly ties over there.” 
On March 19, 2014, Milonov reportedly made anti-Semitic statements to St. Petersburg legislative council. According to the svodka.net news website, Milonov stated that Jews "vilify any saint, it is in their tradition of 2,000 years, beginning with the appeals to crucify the Savior, ending with accusations of anti-Semitism against St. John of Kronstadt." Regarding allegations that St. John of Kronstadt, a 19th-century religious leader, was a supporter of the Black Hundred, Milonov argued that this allegation was based on “complete lies, a modern neo-liberal fable with a sulfuric, deep history of Satanism."
In 2015, Milonov reported that he proposed to the Minister of Internal Affairs to introduce compulsory driving license for bicycle riders, since they behave improperly on the road.
Milonov was married to Eva Liburkina between 2008 and 2011, but is now divorced. They raised three children —daughter Marfa, son Nikolay and a foster-son. Milonov is well known as a deeply religious Christian. He was demonstratively wearing a shirt with slogan "Orthodoxy or death!".
- Молодежные движения и субкультуры Санкт-Петербурга V. V. Kosti︠u︡shev, Institut sot︠s︡iologii (Rossiĭskai︠a︡ akademii︠a︡ nauk). Sankt-Peterburgskiĭ filial, Sankt-Peterburgskiĭ gosudarstvennyĭ universitet. Fakulʹtet sot︠s︡iologii - 1999 p115 "Виталий Милонов практически полностью раскрыл в нем основные идеи, которые исповедуют Молодые Христианские Демократы Санкт-Петербурга. Что касается самой фигуры лидера и отношения к нему других членов МХД, ..."
- St. Petersburg Times Milonov: 'The Sin of Sodom Is Repellent To Me' 2012 3 28 "Vitaly Milonov, the man behind the controversial bill against “promoting sodomy, lesbianism, bisexualism and transgenderism to minors,” fully intends to catch Madonna committing a possible offense when the pop diva visits ..."
- Schwirtz, Michael. Anti-Gay Law Stirs Fears in Russia, The New York Times, 29 Feb 2012
- "Vitaly Milonov will become Duma member" (in Russian). Interfax. 19 September 2016.
- Radia, Kirit, Russian lawmaker: Gay athletes could be prosecuted at 2014 games, WHAS11, 1 Aug 2013, http://www.whas11.com/news/world/Gay-athletes-could-be-prosecuted-at-2014-Russian-games-217919921.html
- Chernov, Serge (2013-11-27), "LGBT Film Festival Faces Bomb Threats and Cancellations", St. Petersburg Times
- "Russian Official Says Apple CEO Should Be Banned From Russia After Saying He Is Gay". Retrieved 20 September 2016.
- "Russia's Deadly Campaign". Out. 2014-05-16. Retrieved 2015-04-08.
- Putin ally accused of pushing Jews-killed-Jesus trope, Jewish Telegraphic Agency, April 2, 2014.
- Russian Jews fear anti-Semitism amid Crimea fervor by Stephen Ennis, BBC, March 28, 2014.
- Milonov, Vitaly (29 June 2015). "Пора вводить права для велосипедистов" (PDF) (in Russian). Smena. p. 6. Retrieved 30 June 2015.
- "Милонов попросил Медведева переименовать Стамбул в Константинополь" (in Russian). RIA Novosti. 15 June 2016. Retrieved 15 June 2016.
- "В начале сентября депутат Виталий Милонов усыновил мальчика". Retrieved 20 September 2016.
- Deputy Milonov about the slogan "Orthodoxy or death!" on his shirt: this is not extremism
- "Замечательный рандом". Retrieved 20 September 2016.
- "Игровые новости > "Большой турнир" Hearthstone в эфире "Игромании"!". Retrieved 20 September 2016.