The invention discloses a preparation method and application of an afterfinishing auxiliary agent to in vitro curly hair. The auxiliary agent consists of alkyl quaternary ammonium salt or fatty amine or an amine salt type cationic surfactant, an emulsifying agent, a silicone elastomer, amino silicone oil, modified silicone, an adhesive moisture-preserving shaping agent, and deionized water. The preparation method comprises the following steps: the cationic surfactant, an emulsifying component and water are stirred and mixed evenly in an emulsifying machine at a temperature of between 70 and 90 DEG C to obtain a transparent and uniform liquid, and the temperature is reduced to 60 DEG C, the pH value is adjusted to be weakly acidic; and then other compositions are added into the liquid, and the mixture is stirred at constant temperature for 0.5 to 1.5 hours until the mixture is mixed evenly and then is cooled and neutralized. When the auxiliary agent is used, the auxiliary agent and softened water are fully dissolved, and fiber to be treated is soaked and then is fished out to be dehydrated and dried by a centrifugal machine. The auxiliary agent has good functions of absorption film-forming and moisture preservation, and can endow curly hair fiber with good sensory characteristics such as softness, smoothness, brightness, high elasticity, fouling prevention, antistatic electricity and so on.