While concert-goers formed a long queue in Gangneung, which is hosting Olympic events, protesters across the street carried placards reading ""
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Others waved a distorted image of North Korean leader Kim Jong Un photoshopped to resemble a demonic boar.
Some 140 members of North Korea's Samjiyon Orchestra were giving their first performance at the Gangneung Art Centre since arriving in the South Tuesday.
The concert takes place a day ahead of the Pyeongchang Olympics opening ceremony, where the two Koreas will march together under the blue-and-white Korean unification flag.
"The concert must be stopped," said Park Byung-seung, who had travelled from Seoul for the protest. "That's promoting Kim Jong Un and North Korea's ideology."
Another protester, with a giant South Korean flag wrapped around her shoulders, said: "Don't you know what is at stake? It's our Olympics and we can't wave the South Korean flag."
Protesters say North Korea, which only confirmed its participation last month, has been allowed to hijack the Pyeongchang Games.
The nuclear-armed North is on an Olympics-linked charm offensive -- sending a troupe of performers, hundreds of female cheerleaders, the ceremonial head of state and even Kim's sister to South Korea.
However, North Korea also held a massive military parade in Pyongyang on Thursday, raising suspicions in the South over the sudden rapprochement, which follows a series of weapons tests.
"By holding the military parade a day before the Olympics and showing off its military power made me doubt Kim Jong Un's intentions," said Lee Young-wook, a protester from Seoul.
Despite the misgivings of some South Koreans, tickets for the orchestra's concerts have been in high demand, with 156,000 people applying for 530 pairs of available seats.
The Koreas' separation makes citizens of the North an object of some fascination for Southerners.
"Not everyone gets to see it," said Kim Mi-sook, who came to the concert with her husband.
"I think it will be a once-in-a-lifetime opportunity unless we are reunified," she added.
The band were expected to perform a mix of South Korean pop songs from the 1980s, as well as North Korean music.
With less than two hours before the performance, North and South Korean officials wrestled over the play list, as some of its own songs had controversial lyrics.
One of the problematic songs was "Moranbong", whose lyrics praise Pyongyang as "the capital of revolution".
The Samjiyon Orchestra will leave for Seoul after the concert to prepare for their second performance on Sunday.
North Korea will have 22 athletes competing at the Pyeongchang Games, but the total number of North Koreans crossing the border is more than 500.
It will be the first time North Korea has attended an Olympics in the South, after they boycotted the 1988 Summer Games in Seoul.
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