Work is a major contributor to our health and well-being. Workers’ thriving is directly influenced by their job design, work environment and organization. The purpose of this report is to describe the qualitative methods used to develop the candidate items for a novel measure of Thriving from Work through a multi-step iterative process including: a literature review, workshop, interviews with experts, and cognitive testing of the candidate items. Through this process, we defined Thriving from Work as the state of positive mental, physical, and social functioning in which workers’ experiences of their work and working conditions enable them to thrive in their overall lives, contributing to their ability to achieve their full potential in their work, home, and community. Thriving from Work was conceptualized into 37 attributes across seven dimensions: psychological, emotional, social, work–life integration, basic needs, experience of work, and health. We ultimately identified, developed and/or modified 87 candidate questionnaire items mapped to these attributes that performed well in cognitive testing in demographically and occupationally diverse workers. The Thriving from Work Questionnaire will be subjected to psychometric testing and item reduction in future studies. Individual items demonstrated face validity and good cognitive response properties and may be used independently from the questionnaire.