The trio are:
DigitalBridge, a tool that uses computer vision and machine learning to let consumers visualise home furnishings;
Link Big, a tool that allows Instagram users to buy products directly from their feed;
Wedding Planner, a service that combines technology and industry expertise to offer wedding planning on mobile and online.
DigitalBridge will receive an investment of £100,000, while the other two will get £50,000 each. All three companies also receive contracts to work with John Lewis.
Two other companies participated in the JLAB programme, but were not chosen to receive funding. One of these, however, Ding Labs, launched a Kickstarter campaign to help it go into full production earlier this month, met its $50,000 (£40,900) target within a week and has now raised more than $95,000. Ding creates a smart doorbell that allows homeowners to talk to the person at their door from anywhere in the world.
The fifth is Robotical, the creators of Marty the Robot, a programmable walking droid that helps kids to learn about coding. That project was also successfully crowdfunded in August on Indiegogo.
John Lewis Partnership chief information officer Paul Coby said: "JLAB was inspired by our founder Spedan Lewis' ideas about bringing innovation to the retail industry.
"The three startups we have invested in have the potential and the technology to really excite John Lewis shoppers."