Running for over 231.23 miles (372.13 km), the segment of I-85 is the longest of the five states it passes through and the second-longest Interstate Highway in North Carolina after I-40.

Landscapes along the route include rolling hills and gently sloping terrain of the Appalachian Mountains at its southernmost stretch, the urbanized neighborhoods of the Piedmont cities through the center of North Carolina, and flat farmlands in the northeast towards Virginia.

I-85 is maintained by the North Carolina Department of Transportation (NCDOT) for its entire length in the state and designated as a Blue Star Memorial Highway.

[5] In 2023, the busiest stretch along the Interstate was from the Statesville Avenue interchange to Graham Street in Charlotte, which carried a total of 200,529 vehicles per day.

A couple of miles later, I-85 has an exit for the airport via Little Rock Road at a single-point urban interchange (SPUI) and enters the city of Charlotte.

[30] It then traverses the northern part of Charlotte, and the routing through this portion, while it does not serve the central business district of the city directly, does pass through largely suburban and urbanized areas near Uptown.

[40] Passing through a mix of suburban and rural development, the highway approaches Salisbury and US 601 splits from I-85 at exit 75 for Jake Alexander Boulevard.

[45][46] After the interchange, I-85 passes underneath the bridge, turning slightly east for one mile (1.6 km), and intersects NC 47 (Hargrave Road).

[7][51] The reversed lanes of I-85 then pass over Squire Bowers Road and reach a rest area, as well as access to the North Carolina Vietnam Veterans Memorial Park.

[59] US 29 forms a brief concurrency with I-85 before the route enters suburban areas once more and reaches a very large and complex interchange with Groometown Road, Grandover Parkway, I-73, US 220, and US 421.

Leaving the massive interchange, I-85 follows the southern outskirts of Greensboro, with US 421 staying connected for just four miles (6.4 km), before US 421 departs the concurrency at exit 126 to head southeast for Sanford.

Meanwhile, I-85 maintains its northeastward track and passes by a couple more exits before reaching I-785 (its third auxiliary route), I-40, and I-840, the former and latter of which have their southern and eastern terminus at I-85 respectively.

[62][63] Past a diverging diamond interchange with NC 119 (Mebane-Oaks Road), the highway enters Orange County and reaches another truck weigh station.

[64] The following interchanges of I-85 before Durham County are rather substandard in quality due to the Interstate retaining its original narrow design of four lanes.

The highway passes a diamond interchange with NC 157 (Guess Road), and then US 501 splits off at Duke Street to head north.

[7][79] Just before leaving North Carolina, I-85 has its final interchange in the state with US 1 and the northern terminus of US 401 near the unincorporated community of Wise, with the southbound lanes then having an exit for the welcome center.

[108] Beginning in what was then known as Fort Henry and now is Petersburg, the travelers made their way into the state through the land now part of Granville County.

[109] In 1799, when William Whedbee Kirkland built his house, Ayr Mount, the Great Trading Path had become a major thoroughfare through the state, with lots of cargo being transported.

[112] One planned road was the Salisbury Bypass, 15 miles (24 km) long with a $1-million (equivalent to $8.88 million in 2023[113]) 880-foot (270 m) twin-span bridge over the Yadkin River.

Green guide signs were also erected to warn of any upcoming access roads ahead in case travelers would want to exit off the Interstate.

To mitigate stress and panic, the common I-85 interstate shield with the colors red, white, and blue would occasionally show up to keep drivers relaxed, calm, and figuring out which way they were going.

This link of I-85 would allow drivers to bypass Franklin Boulevard, the main street through the city, which was often congested and avoid over 22 traffic lights along the road.

[122] The stretch of I-85 from Henderson to the Virginia state line, a distance of approximately 3.1 miles (5.0 km) opened to traffic on December 2, 1970, with a cost of $2 million.

[123] On December 15, 1970, governor Bob Scott announced that about a year from then, all of I-85 would be completed in the state except for a stretch between Greensboro and Lexington.

Costing approximately $16 million, this stretch of the highway was often nicknamed "the missing link", considered as one of the deadliest roads in the state.

This also involved upgrading 94 traffic lights, adding two signs, and placing 11 cameras into the system to help predict any potential accident.

[137][138] In 2024, from the US 321 interchange to I-485 through Gaston and Mecklenburg counties, due to congestion along the Interstate, the lanes are being widened to accommodate larger amounts of traffic.

[144] A study by GPS tracking company Teletrac determined that I-85 in North Carolina was one of the deadliest and most dangerous roads to travel on in Charlotte, ranking 15th out of 30.

While the crash did not cause any damage to I-85 itself, the southbound lanes were closed for several days to repair and maintain the bridge while directing traffic along the interchange ramps.

[159][160] I-785 serves as a spur route, forming a portion of the eastern part of the Greensboro Urban Loop and in the future will connect to Danville, Virginia.