After leaving university, Neville-Rolfe worked in the Civil Service. She worked at the Ministry for Agriculture, Fisheries and Food from 1973–1992. She was a Member of the Prime Minister's Policy Unit from 1992 to 1994. She served as Director of the Deregulation Unit in the Cabinet Office from 1995–97. Neville-Rolfe then took up a position at Tesco and served as Group Director of Corporate Affairs from 1997 to 2006. She served as Company Secretary from 2004–2006. She served on the Board from 2006 as Executive Director until she retired in January 2013. While at Tesco the company moved from its core UK grocery roots into non-food services – and 13 overseas markets across the world. National Life Stories conducted an oral history interview with Neville-Rolfe in 2005-2008 for its Tesco: An Oral History collection held by the British Library. Neville-Rolfe joined the House of Lords as a Conservative Peer in October 2013 and served as Parliamentary Under Secretary of State at the Department for Business, Innovation and Skills and Minister for Intellectual Property from July 2014 until July 2016. From May 2015 she was also Parliamentary Under Secretary of State at the Department for Culture, Media and Sport. Neville-Rolfe was appointed Minister of State at the Department for Business, Energy and Industrial Strategy on 17 July 2016. Before assuming her ministerial responsibilities she spoke in the House of Lords on business, vocational education, broadband, regulatory reform and competitiveness issues. Neville-Rolfe is a particularly prominent speaker on issues concerning business. The aim is to encourage government to facilitate and support UK businesses, to remove unnecessary tax and regulatory burdens, to roll-out broadband across the UK and to enable the growth of small businesses. She has also delivered speeches on UK foreign trade agreements, with particular interest in China and India. She also sat on the Parliamentary All-Party Parliamentary Group for Affordable Childcare. Neville-Rolfe had many non-executive positions which she resigned on appointment. She was a non-executive director of ITV Plc and a member of the supervisory board of Metro Group, a large German-based international retailer and wholesaler. Neville-Rolfe was also President of EuroCommerce, the pan-European retail trade association, and sat on the boards of 2 Sisters Food Group and Hermes Equity Ownership Services and on PwC's Advisory Board. She is a member of the London Business School's Governing Body.
Neville-Rolfe is married to Sir Richard Packer, who was Permanent Secretary at Ministry of Agriculture, Fisheries and Food from 1993 until 2000. They have four sons. Her husband was knighted in 2001. From 2001 until she entered the House of Lords in 2013, her title was Lady Packer.