The John Lewis Partnership will not reopen its 38,000 sq ft At Home store at Greyhound Retail Park as it looks to cut costs amid changing consumer habits.
As well as its Chester shop, which opened in 2011, At Home stores in Ashford, Basingstoke, and Tunbridge Wells will also close.
Department stores in Aberdeen, Peterborough, Sheffield, and York have also traded for the last time.
John Lewis’ other 34 stores will reopen on 12 April when lockdown restrictions allow, the company said.
The partnership also plans to transfer the running of its Waitrose Distribution Centre in Leyland, Lancashire, to XPO Logistics.
John Lewis’ shift in strategy follows the publication of the company’s financial results earlier this month that revealed the firm had made a full-year loss for the first time in its history.
Sharon White, chairman of the John Lewis Partnership, said: “The high street is going through its biggest change for a generation and we are changing with it. Customers will still be able to get the trusted service that we are known for, however and wherever they want to shop.”
John Lewis stores that have survived the cull elsewhere in the North West include the those at Trafford Centre, Cheadle, and Liverpool.